Methods and Composition for Increasing Muscle Protein Synthesis and/or Functional Strength in Mammals

ABSTRACT

A method and composition for preserving muscle mass and function by increasing muscle protein synthesis and/or decreasing muscle protein degradation in mammals is disclosed. In one embodiment, the mammals are administered a protein building composition comprising at least two of an essential amino acid, an amino acid derivative, and a nitrogenous organic acid. In a particular embodiment, the protein building composition comprises leucine, L-carnitine, and creatine. The protein building composition can decrease TNF-α and increase mTOR expression in muscle.

RELATED APPLICATIONS

This application is a continuing application from U.S. patentapplication Ser. No. 15/381,731, having a filing date of Dec. 16, 2016,which claims priority to U.S. Provisional Patent Application No.62/273,880 having a filing date of Dec. 31, 2015, and U.S. ProvisionalPatent Application No. 62/269,573 having a filing date of Dec. 18, 2015,which are incorporated herein in their entirety by reference.

BACKGROUND

Many mammals may experience functional deterioration as they enter midto late adulthood. Functional deterioration relates to a decline in theability to carry out basic functional activities such as eating andwalking. Compromised motor function can, in turn, lead to loss ofindependence and a decreased quality of life.

A major contributor to functional deterioration in both sedentary andphysically active mammals is sarcopenia, an age-related conditioncharacterized by a gradual decline in muscle mass, strength, andfunction. In sarcopenia, the body experiences a global shift from muscleprotein synthesis to muscle degradation.

Proteinogenic amino acid molecules can be categorized as essential andnon-essential amino acids. Non-essential amino acids, which can besynthesized in the body, include alanine, asparagine, aspartic acid,glutamic acid, arginine, cysteine, glutamine, tyrosine, glycine,proline, and serine. Essential amino acids, such as histidine,isoleucine, leucine, lysine, methionine, phenylalanine, threonine,tryptophan, and valine, cannot be biosynthesized and instead must beconsumed via food or supplementation. In muscle protein synthesis, theavailability of essential amino acids is rate-limiting. Therefore,supplementation of essential amino acids can potentiate proteinsynthesis. Muscle protein synthesis is activated by the mammalian targetof rapamycin (mTOR) pathway. The mTOR pathway senses and responds tochanges in amino acids and growth factors such as insulin-like growthfactor (IGF-1). In particular, the mTOR complex phosphorylates ribosomalprotein S6K, which in turn phosphorylates ribosomal protein S6 andsubstrates eIF4B and 4EBP1, consequently promoting mRNA translation andprotein synthesis. The proteins involved in the mTOR pathway can serveas biomarkers for protein synthesis.

The NFκβ (p65/p50) pro-inflammatory signaling pathway has also beenimplicated in regulating muscle functionality. NFκβ protein levels arestrongly upregulated in muscle atrophy, while inhibition of the NFκβ(p65/p50) pathway can prevent muscle degradation.

The ubiquitin-proteasome system (UPS) can facilitate muscle degradation,breaking down muscle proteins into peptides and then free amino acids.In the UPS-pathway, target muscle proteins are tagged with a degradationsignal and then degraded into peptides by the 26s proteasome. TheUPS-pathway can be mediated by atrogin-1 and MuRF-1, two E3ubiquitin-protein ligases, and by cytokines such as interleukin (IL)-6,IL-8, and tumor necrosis factor (TNF)-α, which produce pro-inflammatorysignals. These factors can serve as biomarkers for muscle proteindegradation. Protein degradation and functional deterioration may beamplified in mammals with sarcopenia and in older mammals with sedentarylifestyles. Muscle protein building requires both new protein synthesisand prevention of protein degradation. While physical activity, even inlight or moderate amounts, can lead to muscle hypertrophy and increasedfunctionality, muscle disuse results in a decrease in protein synthesisand an increase in muscle breakdown. In addition, mTOR activation can bedelayed in older adults, leading to decreased protein synthesis.Furthermore, extended periods of inactivity can lead to a decreasedprotein synthetic response to amino acid supplementation.Sarcopenia-related functional deterioration can be a reinforcing loop,as muscle loss and decreased functionality can interfere with physicalactivity. In turn, decreased physical activity leads to further muscleloss and decreased functionality.

In the past, leucine and creatine have been administered to mammals inorder to increase muscle mass and strength. In particular, many of thesupplements administered to elderly mammals for increasing muscleprotein synthesis comprise complicated and expensive mixtures of aminoacids. For example, U.S. Pat. No. 7,790,688, which is incorporated byreference herein, discloses compositions containing a complex blend ofessential amino acids, creatine, and low-glycemic carbohydrates.Further, it is well-known in the art that ingestion of 20 grams or moreof essential amino acids is necessary for stimulation of muscle proteinsynthesis in the elderly. As protein intake increases, however, ureaproduction can increase in some individuals.

Consequently, a need exists for a supplement that can increase muscleprotein synthesis, lean mass, functional strength, and/or overallquality of life in mammals without having any substantial adverseeffects on other body functions. A need also exists for a simpler andless expensive supplement that can improve muscle mass and/or functionalstrength leading to increase in physical activity in elderly mammals andmammals with sarcopenia.

SUMMARY

The present disclosure is generally directed to a method and compositionfor increasing protein synthesis and/or functional strength in mammals.The present disclosure is also directed to a method and composition forslowing down or delaying muscle wasting or sarcopenia in sedentary andphysically active mammals.

In accordance with the present disclosure, the method comprises the stepof administering to a mammal an effective amount of a protein buildingcomposition. Of particular advantage, the protein building compositioncan increase muscle protein synthesis, lean mass, functional strength,physical activity, and/or overall quality of life in mammals withouthaving any substantial adverse effects on other body functions.

The protein building composition may comprise a supplement, a foodproduct, or a beverage. For instance, the protein building compositioncan be incorporated into milkshake drinks, juices, cereal bars, vitaminsincluding gummy vitamins, powders, foods or may be in the form of asupplement.

In one embodiment, the protein building composition may be administeredto a mammal that has an age of more than 50% of its expected life span.In an alternate embodiment, the protein building composition may beadministered to a mammal that has an age of less than 50% of itsexpected life span. The mammal may regularly participate in physicalactivity. In another embodiment, the mammal may participate mainly orexclusively in sedentary behavior. In a particular embodiment, themammal is a human.

In one embodiment, the protein building composition may comprise atleast two of any essential amino acid components, amino acidderivatives, and/or nitrogenous organic acids, such as creatine. Theessential amino acid component may comprise leucine and derivativesand/or salts thereof. The amino acid derivative may comprise carnitineand derivatives and/or salts thereof. In one embodiment, the proteinbuilding composition comprises an amino acid derivative combined with anamino acid component. In an alternative embodiment, the protein buildingcomposition may comprise an amino acid derivative combined with anitrogenous organic acid. In still another embodiment, the proteinbuilding composition may comprise a mixture of an amino acid derivative,an amino acid component, and a nitrogenous organic acid.

In one embodiment, the amino acid derivative comprises L-carnitine. In aparticular embodiment, the amino acid derivative comprises L-carnitineL-tartrate. In a further embodiment, the protein building compositionmay contain a nitrogenous organic acid and derivatives and/or saltsthereof. In one embodiment, the protein building composition maycomprise creatine and derivatives and/or salts thereof.

In one embodiment, leucine and salts, metabolites, and/or derivativesthereof may be present in the amino acid component at a concentrationgreater than about 5% by mass. In one embodiment, a metabolite ofleucine, such as β-Hydroxy β-methylbutyric acid (HMB), may be present inthe amino acid component. In one embodiment, leucine is present in aconcentration of about 10% to 90% by mass, such as about 20% to 80% bymass, such as about 30% to 70% by mass. In a preferred embodiment,leucine is present in a concentration of about 35 to 65% by mass.

In one embodiment, L-carnitine may be present in the amino acidcomponent at a concentration greater than about 5% by mass. L-carnitinemay be present in a substantially pure crystalline form or as salts,metabolites, and/or lipid and non-lipid derivatives of L-carnitine. Inone embodiment, L-carnitine is present in a concentration of about 10%to 90% by mass, such as about 20% to 80% by mass, such as about 25% to75% by mass, such as about 30% to 60% by mass. In a preferredembodiment, L-carnitine is present in a concentration of about 40% to70% by mass.

In one embodiment, creatine and salts, metabolites, and/or derivativesthereof may be present in the supplement composition at a concentrationof greater than about 0.5% by mass, such as greater than about 1% bymass, such as greater than about 5% by mass, such as greater than about10% by mass, such as greater than about 20% by mass, such as greaterthan about 40% by mass, such as greater than about 60% by mass, such asgreater than about 80% by mass, such as greater than about 90% by mass,such as greater than about 99% by mass. In a preferred embodiment,creatine is present in the composition at a concentration of about 30%to about 45% by mass. In one embodiment, creatine may comprise creatinemonohydrate and chelated creatine, such as magnesium-chelated creatineand other metal-chelated creatine. The protein building composition canbe administered to the mammal regularly or occasionally. For instance,the protein building composition can be administered to the mammal atleast every one to three days, such as daily. In one embodiment, a dailyportion may be taken in one or several servings. Each dose may be fromabout 5 milligrams to about 30,000 milligrams, such as from about 5milligrams to about 20,000 milligrams, such as from about 50 milligramsto about 10,000 milligrams. Each dose may be from about 1 milligram perkilogram body weight per day to about 10,000 milligrams per kilogram perday, such as from about 5 milligrams per kilogram per day to about 5,000milligrams per kilogram per day. The protein building composition can beadministered orally and can be combined with a food composition.

In one embodiment, the pharmaceutical composition of the presentdisclosure is combined with various additives and ingredients in orderto improve various properties. For instance, the protein buildingcomposition of the present disclosure may be combined with a stabilizerpackage for reducing the hydroscopic properties of the composition. Thestabilizer package can also make the composition easier to handle and/orpour, especially when the composition comprises a granular composition.

In one embodiment, for instance, the present disclosure is directed to apharmaceutical composition comprising a protein building compositioncombined with a polymer binder and a stabilizer package. The stabilizerpackage may comprise oxide particles, such as silica, combined with asalt of a carboxylic acid. The salt of a carboxylic acid may comprise asalt of a fatty acid, such as a fatty acid having a carbon chain lengthof from about 6 carbon atoms to about 40 carbon atoms, such as fromabout 12 carbon atoms to about 28 carbon atoms. In one embodiment, thesalt of the carboxylic acid comprises a stearate salt, such as calciumstearate, sodium stearate, magnesium stearate, mixtures thereof, and thelike. The polymer binder, on the other hand, may comprise apolysaccharide and/or a film-forming polymer. The polymer binder, forinstance, may comprise starch such as a modified starch, maltodextrin,gum arabic, arabinogalactan, a gelatin, or mixtures thereof.

In one embodiment, the pharmaceutical composition may further contain acoating material. The coating material may comprise a fat. The coatingmaterial may form a continuous or a discontinuous coating over thepharmaceutical composition. In one embodiment, the coating materialcomprises a hydrogenated palm oil combined with palm stearine. Forinstance, the hydrogenated palm oil and palm stearine may be combined ata weight ratio of from about 10:1 to about 1:1, such as from about 5:1to about 3:1.

The present disclosure is also directed to a method of producing apharmaceutical composition containing the protein building composition.The method includes the steps of combining the protein buildingcomposition with a polymer binder and a stabilizer package. In oneembodiment, for instance, the protein building composition is firstcombined with the polymer binder via a spray dry process and thencombined with the stabilizer package which may comprise a dry mix (i.e.a powder or granular material). The method may further include the stepof optionally applying a coating material to the mixture containing theprotein building composition, the polymer binder, and the stabilizerpackage. The coating material may comprise a fat, such as a hydrogenatedoil.

In one embodiment, the pharmaceutical composition contains the oxideparticles, such as the silica particles, in an amount from about 0.01%to about 1.5% by weight, the salt of the carboxylic acid in an amountfrom about 0.5% to about 5% by weight and the polymer binder in anamount from about 8% to about 40% by weight. When present, the coatingmaterial may be contained in the polymer composition in an amount fromabout 5% to about 35% by weight.

Other features and aspects of the present disclosure are discussed ingreater detail below.

BRIEF DESCRIPTION OF THE DRAWINGS

A full and enabling disclosure of the present disclosure is set forthmore particularly in the remainder of the specification, includingreference to the accompanying figures, in which:

FIG. 1 shows results of the change in absolute total lean mass frombaseline to week 8 for participants supplemented with either placebo,Carnipure product 1, or Carnipure product 2 in the PP Population (N=39),obtained by the procedure described in Example 1;

FIG. 2 shows results of the change in arm lean mass from baseline toweek 8 for participants supplemented with either placebo, Carnipureproduct 1, or Carnipure product 2 in the PP Population (N=39) obtainedby the procedure described in Example 1;

FIG. 3 shows results of the change in leg lean mass from baseline toweek 8 for participants supplemented with either placebo, Carnipureproduct 1, or Carnipure product 2 in the PP Population (N=39) obtainedby the procedure described in Example 1;

FIG. 4 shows results of the change in total non-trunk lean mass frombaseline to week 8 for participants supplemented with either placebo,Carnipure product 1, or Carnipure product 2 in the PP Population (N=39)obtained by the procedure described in Example 1;

FIG. 5 shows results of the change in absolute average leg strength frombaseline to week 8 for participants supplemented with either placebo,Carnipure product 1, or Carnipure product 2 in the PP Population (N=39)obtained by the procedure described in Example 1;

FIG. 6 shows results of the AUC per μg of protein for Phospho-mTOR frombaseline to week 8 pre and post exercise for participants supplementedwith either placebo, Carnipure product 1, or Carnipure product 2 in thePP Population (N=33) obtained by the procedure described in Example 1;

FIG. 7 shows results of the change in Phospho-mTOR from baseline to week8 pre and post exercise for participants supplemented with eitherplacebo, Carnipure product 1, or Carnipure product 2 in the PPPopulation (N=33) obtained by the procedure described in Example 1;

FIG. 8 shows results of the AUC per μg of protein for Total-mTOR frombaseline to week 8 pre and post exercise for participants supplementedwith either placebo, Carnipure product 1, or Carnipure product 2 in thePP Population (N=33) obtained by the procedure described in Example 1;

FIG. 9 shows results of the change in Total-mTOR from baseline to week 8pre and post exercise for participants supplemented with either placebo,Carnipure product 1, or Carnipure product 2 in the PP Population (N=33)obtained by the procedure described in Example 1;

FIG. 10 shows results of the AUC per μg of protein for Phospho-S6K frombaseline to week 8 pre and post exercise for participants supplementedwith either placebo, Carnipure product 1, or Carnipure product 2 in thePP Population (N=33) obtained by the procedure described in Example 1;

FIG. 11 shows results of the Change in Phospho-S6K from baseline to week8 pre and post exercise for participants supplemented with eitherplacebo, Carnipure product 1, or Carnipure product 2 in the PPPopulation (N=33) obtained by the procedure described in Example 1;

FIG. 12 shows results of the AUC per μg of protein for Total-S6K frombaseline to week 8 pre and post exercise for participants supplementedwith either placebo, Carnipure product 1, or Carnipure product 2 in thePP Population (N=33) obtained by the procedure described in Example 1;

FIG. 13 shows results of the Change in Total-S6K from baseline to week 8pre and post exercise for participants supplemented with either placebo,Carnipure product 1, or Carnipure product 2 in the PP Population (N=33)obtained by the procedure described in Example 1;

FIG. 14 shows results of the AUC per μg of protein for Phospho-4EBP1from baseline to week 8 pre and post exercise for participantssupplemented with either placebo, Carnipure product 1, or Carnipureproduct 2 in the PP Population (N=33) obtained by the proceduredescribed in Example 1;

FIG. 15 shows results of the change in Phospho-4EBP1 from baseline toweek 8 pre and post exercise for participants supplemented with eitherplacebo, Carnipure product 1, or Carnipure product 2 in the PPPopulation (N=33) obtained by the procedure described in Example 1;

FIG. 16 shows results of the AUC per μg of protein for Total-4EBP1 frombaseline to week 8 pre and post exercise for participants supplementedwith either placebo, Carnipure product 1, or Carnipure product 2 in thePP Population (N=33) obtained by the procedure described in Example 1;

FIG. 17 shows results of the change in total-4EBP1 from baseline to week8 pre and post exercise for participants supplemented with eitherplacebo, Carnipure product 1, or Carnipure product 2 in the PPPopulation (N=33) obtained by the procedure described in Example 1;

FIG. 18 shows results of the AUC per μg of protein for Phospho-P65 frombaseline to week 8 pre and post exercise for participants supplementedwith either placebo, Carnipure product 1, or Carnipure product 2 in thePP Population (N=33) obtained by the procedure described in Example 1;

FIG. 19 shows results of the change in Phospho-P65 from baseline to week8 pre and post exercise for participants supplemented with eitherplacebo, Carnipure product 1, or Carnipure product 2 in the PPPopulation (N=33) obtained by the procedure described in Example 1;

FIG. 20 shows results of the AUC per μg of protein for Total-P65 frombaseline to week 8 pre and post exercise for participants supplementedwith either placebo, Carnipure product 1, or Carnipure product 2 in thePP Population (N=33) obtained by the procedure described in Example 1;

FIG. 21 shows results of the change in Total-P65 from baseline to week 8pre and post exercise for participants supplemented with either placebo,Carnipure product 1, or Carnipure product 2 in the PP Population (N=33)obtained by the procedure described in Example 1;

FIG. 22 shows results of the AUC per μg of protein for Total-P50 frombaseline to week 8 pre and post exercise for participants supplementedwith either placebo, Carnipure product 1, or Carnipure product 2 in thePP Population (N=33) obtained by the procedure described in Example 1;

FIG. 23 shows results of the change in Total-P50 from baseline to week 8pre and post exercise for participants supplemented with either placebo,Carnipure product 1, or Carnipure product 2 in the PP Population (N=33)obtained by the procedure described in Example 1;

FIG. 24 shows results of a representative immune-blot shown frombaseline to week 8 for participants supplemented with either placebo,Carnipure product 1, or Carnipure product 2 in the PP Population (N=33)obtained by the procedure described in Example 1; and

FIG. 25 shows results of the change in absolute composition score frombaseline to week 8 for participants supplemented with either placebo,Carnipure product 1, or Carnipure product 2 in the PP Population (N=39)obtained by the procedure described in Example 1.

The Figures are graphical representations of the results obtained in theexample described below.

DEFINITIONS

The term “MET,” or “metabolic equivalent,” means the ratio of the rateof energy expended during an activity to the rate of energy expended atrest. A body at rest has a rate of energy expenditure of 1 MET. If abody performs a 2 MET activity, the body has expended 2 times the energyused by the body at rest. The term “physical activity” means bodilymovement with an energy expenditure rate equal to or greater than 3 MET.Non-limiting examples of “physical activity” include bicycling, sexualactivity, giving birth, jogging, walking at a speed of about 3 mph orgreater, calisthenics, jumping rope, running, sprinting, or anycombinations thereof.

In one embodiment, “physical activity” can mean a negative energybalance in the mammal, such as weight loss, diets, aging, gestation, andlactation.

The term “physically active” means regularly participating in bodymovements with an energy expenditure rate of greater than or equal to 3MET.

In one embodiment, “physically active” can mean regularly meetingmedically recommended standards for amount, intensity, and type ofphysical activity performed by a mammal.

The terms “sedentary” or “sedentary activity” mean participating mainlyor exclusively in body movements with an energy expenditure rate of lessthan 3 MET. Non-limiting examples of “sedentary” activities includesleeping, resting, sitting or reclining, watching television, writing,working at a desk, using a computer, typing, walking at a speed of lessthan about 3 mph, or any combinations thereof.

In one embodiment, “sedentary” can mean a failure to regularly meetmedically recommended standards for amount, intensity, and type ofphysical activity performed by a mammal.

The term “L-carnitine” may contain L-carnitine and derivatives and/orsalts thereof. L-carnitine can include L-carnitine base or derivativesand/or salts thereof including substantially pure crystallineL-carnitine, any fatty acid derivatives thereof, acetyl L-carnitine,valeryl L-carnitine, isovaleryl L-carnitine, benzyl L-carnitine,L-leucyl L-carnitine, L-valyl L-carnitine, other L-amino acylcarnitines, salts of L-amino acyl L-carnitine, L-carnitine HCL,L-carnitine L-tartrate, L-carnitine fumarate, propionyl L-carnitine,L-carnitine phosphate, acetyl L-carnitine L-aspartate, acetylL-carnitine citrate, acetyl L-carnitine maleate, acetyl L-carnitinephosphate, acetyl L-carnitine fumarate, propionyl L-carnitine orotate,acetyl L-carnitine orotate, butyryl L-carnitine orotate, propionylL-carnitine fumarate, L-carnitine oxalate, L-carnitine sulfate, GPLCglycine propionyl L-carnitine, and the like.

The term “mammal” includes any mammal that may experience skeletalmuscle degradation or synthesis and includes human, canine, equine,feline, bovine, ovine, or porcine mammals.

The phrase “effective amount” means an amount of a compound thatpromotes, improves, stimulates, or encourages a response to theparticular condition or disorder or the particular symptom of thecondition or disorder.

The term “supplement” means a product in addition to the normal diet ofthe mammal but may be combined with a mammal's normal food or drinkcomposition. The supplement may be in any form but not limited to asolid, liquid, gel, capsule, or powder.

A supplement may also be administered simultaneously with or as acomponent of a food composition which may comprise a food product, abeverage, a pet food, a snack, or a treat. In one embodiment, thebeverage may be an activity drink.

The term “functional strength” means an individual's ability tocompetently and safely perform daily life activities. In one embodiment,functional strength may be associated with energy, muscle potency,agility, flexibility, balance, and injury resistance.

DETAILED DESCRIPTION

It is to be understood by one of ordinary skill in the art that thepresent discussion is a description of exemplary embodiments only, andis not intended as limiting the broader aspects of the presentdisclosure.

The present disclosure is directed to a method and composition forincreasing muscle protein synthesis and/or functional strength inmammals. As will be explained below, the present disclosure is generallydirected to a protein building composition that when administered to amammal in an effective amount preserves or increases muscle mass andfunction by increasing muscle protein synthesis and/or functionalstrength and/or decreasing protein degradation. The present disclosureis further generally directed to administering to a mammal an effectiveamount of a protein building composition that increases muscle proteinsynthesis and/or functional strength.

In one embodiment, the protein building composition comprises an aminoacid derivative combined with at least one other component. Forinstance, the amino acid derivative may be combined with an amino acid,an organic acid, or both an amino acid and an organic acid. The aminoacid derivate may comprise a carnitine, such as L-carnitine. The aminoacid may comprise an essential amino acid. In one embodiment, the aminoacid may comprise leucine. The protein building composition may furthercomprise an organic acid, such as a nitrogenous organic acid. Theorganic acid may comprise creatine.

The inventors of the present disclosure unexpectedly discovered that thecomponents of the protein building composition disclosed hereinsynergistically work together to increase muscle protein synthesisand/or functional strength. Surprisingly, it was discovered thatadministering a protein building composition containing L-carnitine,combined with leucine, creatine, or both leucine and creatine to amammal can increase muscle protein synthesis and/or functional strengthwhile also preventing muscle degradation, decreasing inflammation, andimproving upper and lower body strength. In one embodiment, the proteinbuilding composition may also contain vitamin D3. Of particularadvantage, the above results can be achieved without administering thehigh dosage of amino acids required in previous compositions. Inaddition, the protein building composition of the present disclosureproduces no noticeable side effects. For instance, administering theprotein building composition regularly can be done without showing anyclinically significant change in kidney function.

The protein building composition of the present disclosure can be anutritional supplement in the form of a pill, tablet or capsule.Alternatively, the protein building composition may be incorporated intoa food or beverage. For instance, the protein building composition maybe incorporated in to a milkshake drink, a juice, a cereal bar, avitamin such as a gummy vitamin, or a powder. The powder can be mix withany suitable liquid for ingestion.

The amino acid derivative may be any suitable carnitine, such asL-carnitine and any derivatives and/or salts thereof. L-carnitine is aquaternary amine that can be biosynthesized from lysine and methionine.L-carnitine is known to promote beta-oxidation of long-chain fatty acidsby facilitating their transfer across the mitochondrial membrane.L-carnitine may be present in a substantially pure crystalline form oras salts, metabolites, and/or lipid and non-lipid derivatives ofL-carnitine.

In one embodiment, L-carnitine may be present in the composition at aconcentration greater than about 5% by mass, such as greater than about10% by mass, such as greater than about 15% by mass, such as greaterthan about 20% by mass, such as greater than about 25% by mass, such asgreater than about 30% by mass, such as greater than about 35% by mass,such as greater than about 40% by mass, such as greater than about 45%by mass, such as greater than about 50% by mass, such as greater thanabout 55% by mass, such as greater than about 60% by mass, such asgreater than about 65% by mass, such as greater than about 70% by mass,such as greater than about 75% by mass. In general, L-carnitine may bepresent in the composition at a concentration less than about 75% bymass, such as less than about 70% by mass, such as less than about 65%by mass, such as less than about 60% by mass, such as less than about55% by mass, such as less than about 50% by mass, such as less thanabout 45% by mass, such as less than about 40% by mass, such as lessthan about 35% by mass, such as less than about 30% by mass, such asless than about 25% by mass, such as less than about 20% by mass, suchas less than about 15% by mass, such as less than about 10% by mass.

As stated above, the composition may comprises an amino acid incombination with the amino acid derivate. The amino acid may be leucineand any derivatives, metabolites, and/or salts thereof. Leucine is abranched-chain α-amino acid. Leucine is known to stimulate proteinsynthesis via association with eukaryotic initiation factors intranslation. In one embodiment, the amino acid may comprise a metaboliteof leucine, such as HMB.

In one embodiment, leucine may be present in the amino acid component ata concentration greater than about 10% by mass, such as greater thanabout 15% by mass, such as greater than about 20% by mass, such asgreater than about 25% by mass, such as greater than about 30% by mass,such as greater than about 35% by mass, such as greater than about 40%by mass, such as greater than about 45% by mass, such as greater thanabout 50% by mass, such as greater than about 55% by mass, such asgreater than about 60% by mass, such as greater than about 65% by mass,such as greater than about 70% by mass, such as greater than about 75%by mass. In general, leucine may be present in the amino acid componentat a concentration less than about 75% by mass, such as less than about70% by mass, such as less than about 65% by mass, such as less thanabout 60% by mass, such as less than about 55% by mass, such as lessthan about 50% by mass, such as less than about 45% by mass, such asless than about 40% by mass, such as less than about 35% by mass, suchas less than about 30% by mass, such as less than about 25% by mass,such as less than about 20% by mass, such as less than about 15% bymass.

As described above, the protein building composition may comprise theamino acid derivative in combination with either an amino acid or anorganic acid or may comprise an amino acid derivative in combinationwith both an amino acid or an organic acid. The organic acid may becreatine and derivatives and/or salts thereof. Creatine is a nitrogenousorganic acid that can be biosynthesized from glycine and arginine.Creatine increases the formation of ATP and in a phosphorylated formserves as an energy reserve in skeletal muscles. Creatine can improvethe physiological response to high-intensity and resistance exercise.

In one embodiment, the composition may include creatine and derivativesand analogs and/or salts thereof. In one embodiment, the composition mayinclude creatine phosphate; creatine monohydrate; creatine ethyl ester;magnesium creatine chelate; creatine HCL; creatine-MG-complex (acetate);phosphocreatine-Mg-complex (acetate); creatine sugar amides and saltsthereof as described in U.S. Pat. No. 8,546,369, incorporated byreference herein; (Boc)2-creatine and derivatives thereof as describedin PCT Publication WO 2014/097335, incorporated by reference herein;other derivatives and salts of creatine; and any combinations thereof.

In one embodiment, creatine may be present in the amino acid componentat a concentration greater than about 15% by mass, such as greater thanabout 20% by mass, such as greater than about 25% by mass, such asgreater than about 30% by mass, such as greater than about 35% by mass,such as greater than about 40% by mass, such as greater than about 45%by mass, such as greater than about 50% by mass, such as greater thanabout 55% by mass, such as greater than about 60% by mass, such asgreater than about 65% by mass, such as greater than about 70% by mass,such as greater than about 75% by mass, such as greater than about 80%by mass. In general, creatine may be present in the amino acid componentat a concentration less than about 80% by mass, such as less than about75% by mass, such as less than about 70% by mass, such as less thanabout 65% by mass, such as less than about 60% by mass, such as lessthan about 55% by mass, such as less than about 50% by mass, such asless than about 45% by mass, such as less than about 40% by mass, suchas less than about 35% by mass, such as less than about 30% by mass,such as less than about 25% by mass, such as less than about 20% bymass, such as less than about 15% by mass, such as less than about 10%by mass.

In one embodiment, the composition may comprise one or more vitamins. Inone particular embodiment, the composition may comprise vitamin D3.Vitamin D can regulate muscle contractility. Other vitamins may includebut are not limited to vitamin A, vitamin B1, vitamin B2, vitamin B3,vitamin B6, vitamin B9, vitamin B12, vitamin C, vitamin E, vitamin K,riboflavin, niacin, folic acid, pyridoxine, thiamine, pantothenic acid,biotin, and any combinations thereof. The composition may furthercomprise other minerals, herbs, botanicals, and essential fatty acids.In one embodiment, the protein building composition may comprisemagnesium and/or salts thereof.

In one embodiment, the one or more vitamins may be present in thecomposition in an amount of about 1 to about 5,000 IU per dose, such asabout 10 to about 2,500 IU per dose, such as about 50 to about 1,500 IUper dose, such as about 100 IU to about 1,000 IU per dose, such as about250 IU to about 750 IU per dose, such as about 300 IU to about 600 IUper dose.

In one embodiment, the amino acids, amino acid derivatives, and/ororganic acids and derivatives and/or salts thereof may be included inthe composition as free form organic compounds. Alternately, thecomponents may be included in the composition as intact proteins and/orother macromolecules. In a further embodiment, the amino acids, aminoacid derivatives, and/or organic acids and derivatives and/or saltsthereof may be included in the composition as a combination of free formorganic compounds and intact protein and/or other macromolecules.

The different components can be present in the protein buildingcomposition at various ratios depending upon the particular applicationand the desired result. The weight ratio between the amino acidderivative and the amino acid, for instance, can generally be from about1:10 to about 10:1, such as from about 1:3 to about 3:1. In oneembodiment, the weight ratio between the amino acid derivative and theamino acid may be from about 1:1 to about 1:2.

The weight ratio between the amino acid derivative and the organic acid,on the other hand, can generally be from about 1:10 to about 5:1, suchas from about 1:1 to about 1:5, such as from about 1:1 to about 1:3. Inone particular embodiment, the protein building composition contains theamino acid derivative combined with the amino acid and the organic acid.The weight ratio between the amino acid derivative and the amino acidcan be about 3:4, while the weight ratio between the amino derivativeand the organic acid can be about 1:2.

The present disclosure is also directed to methods of administering theprotein building composition disclosed herein. L-carnitine incombination with leucine and creatine has been discovered to increaseprotein synthesis and/or functional strength in mammals. In order toincrease muscle protein synthesis and/or functional strength, thepresent disclosure is directed to a method of administering to a mammalan effective amount of a protein building composition. The aboveadvantages and benefits may be realized without any adverseconsequences. In addition, in one embodiment, the mammal may experienceno clinically substantial difference in kidney function.

In one embodiment, a mammal is administered an effective amount of anprotein building composition containing an amino acid component. Theamino acid component may comprise leucine. Leucine may be administeredin a dosage from about 5 to 10,000 milligrams per day, such as fromabout 5 to about 5,000 milligrams per day, such as from about 50milligrams to about 3,000 milligrams per day. The dosage, for instance,can be greater than about 100 milligrams per day, such as greater thanabout 250 milligrams per day, such as greater than about 500 milligramsper day, such as greater than about 750 milligrams per day. Based onbody mass, the dosage can be from about 1 milligram per kilogram of bodyweight per day to about 1,000 milligrams per kilogram body weight perday. For example, the dosage may be from about 5 milligrams per kilogrambody weight per day to about 750 milligrams per kilogram body weight perday. In one particular embodiment, the dosage can be from about 10milligrams per kilogram body weight per day to about 500 milligrams perkilogram body weight per day. In another particular embodiment, thedosage can be greater than about 1 milligrams per kilogram body weightper day, greater than about 5 milligrams per kilogram body weight perday, greater than about 10 milligrams per kilogram body weight per day,greater than about 15 milligrams per kilogram body weight per day,greater than about 20 milligrams per kilogram body weight per day,greater than about 25 milligrams per kilogram body weight per day,greater than about 30 milligrams per kilogram body weight per day, orgreater than about 35 milligrams per kilogram body weight per day.

The mammal can also be administered an effective amount of an amino acidderivative. The amino acid derivative may comprise L-carnitine.L-carnitine may be administered in a dosage from about 5 to 10,000milligrams per day, such as from about 5 to about 5,000 milligrams perday, such as from about 50 milligrams to about 3,000 milligrams per day.The dosage, for instance, can be greater than about 100 milligrams perday, such as greater than about 250 milligrams per day, such as greaterthan about 500 milligrams per day, such as greater than about 750milligrams per day. Based on body mass, the dosage can be from about 1milligram per kilogram of body weight per day to about 1,000 milligramsper kilogram body weight per day. For example, the dosage may be fromabout 5 milligrams per kilogram body weight per day to about 750milligrams per kilogram body weight per day. In one particularembodiment, the dosage can be from about 10 milligrams per kilogram bodyweight per day to about 500 milligrams per kilogram body weight per day.In another particular embodiment, the dosage can be greater than about 1milligrams per kilogram body weight per day, greater than about 5milligrams per kilogram body weight per day, greater than about 10milligrams per kilogram body weight per day, greater than about 15milligrams per kilogram body weight per day, greater than about 20milligrams per kilogram body weight per day, greater than about 24milligrams per kilogram body weight per day, greater than about 28milligrams per kilogram body weight per day, or greater than about 30milligrams per kilogram body weight per day.

In one embodiment, a mammal is administered an effective amount of anprotein building composition containing an organic acid and derivativeand/or salt thereof. The organic acid may comprise creatine. Creatinemay be administered in a dosage from about 5 to 15,000 milligrams perday, such as from about 5 to about 10,000 milligrams per day, such asfrom about 50 milligrams to about 5,000 milligrams per day. The dosage,for instance, can be greater than about 100 milligrams per day, such asgreater than about 500 milligrams per day, such as greater than about1,000 milligrams per day, such as greater than about 1,250 milligramsper day. Based on body mass, the dosage can be from about 1 milligramper kilogram of body weight per day to about 1,000 milligrams perkilogram body weight per day. For example, the dosage may be from about5 milligrams per kilogram body weight per day to about 750 milligramsper kilogram body weight per day. In one particular embodiment, thedosage can be from about 10 milligrams per kilogram body weight per dayto about 500 milligrams per kilogram body weight per day. In anotherparticular embodiment, the dosage can be greater than about 1 milligramsper kilogram body weight per day, greater than about 5 milligrams perkilogram body weight per day, greater than about 10 milligrams perkilogram body weight per day, greater than about 15 milligrams perkilogram body weight per day, greater than about 20 milligrams perkilogram body weight per day, greater than about 30 milligrams perkilogram body weight per day, greater than about 40 milligrams perkilogram body weight per day, or greater than about 50 milligrams perkilogram body weight per day.

The protein building composition can be administered regularly, such asat least two to four times a week. For instance, the protein buildingcomposition may be administered to the mammal at least every one tothree days. Further, the protein building composition may beadministered more than one time per day. For instance, the proteinbuilding composition may be administered to the mammal one to four timesper day. In one particular embodiment, the protein building compositionis administered daily. The dosage can be from about 5 to 30,000milligrams per day, such as from about 5 to about 20,000 milligrams perday, such as from about 10 milligrams to about 10,000 milligrams perday, such as from about 20 milligrams to about 5,000 milligrams per day,such as from about 50 milligrams to about 2,500 milligrams per day.Based on body mass, the dosage can be from about 1 milligram perkilogram of body weight per day to about 10,000 milligrams per kilogrambody weight per day. For example, the dosage may be from about 5milligrams per kilogram body weight per day to about 7,500 milligramsper kilogram body weight per day, such as from about 10 milligrams perkilogram body weight per day to about 5,000 milligrams per kilogram bodyweight per day, such as from about 15 milligrams per kilogram bodyweight per day to about 2,500 milligrams per kilogram body weight perday, such as from about 20 milligrams per kilogram body weight per dayto about 1,000 milligrams per kilogram body weight per day, such as fromabout 25 milligrams per kilogram body weight per day to about 750milligrams per kilogram body weight per day, such as from about 30milligrams per kilogram body weight per day to about 500 milligrams perkilogram body weight per day, such as from about 35 milligrams perkilogram body weight per day to about 250 milligrams per kilogram bodyweight per day.

The protein building composition can be administered to the mammal inany suitable form using any suitable administration route. For example,the composition can be administered orally alone, in combination with afood composition, or as part of a food composition. The composition mayalso be part of a dietary supplement or as a nutraceutical composition.

The protein building composition can be administered orally as a solid,liquid, suspension, or gas. The composition may be administered viabuccal or sublingual administration. In one embodiment, the proteinbuilding composition may be administered as a capsule, tablet, caplet,pill, troche, drop, lozenge, powder, granule, syrup, tea, drink, thinfilm, seed, paste, herb, botanical, and the like.

In addition to being administered orally, the supplement dose can alsobe administered using other routes including intranasal, intravenous,intramuscular, intragastric, and the like.

When the protein building composition is combined with a food orbeverage composition, the food or beverage composition may comprise anysuitable composition for consumption by the mammal. Such compositionsinclude complete foods or beverages intended to supply the necessarydietary requirements for mammal or food supplements such as treats andsnacks. The food composition may comprise pellets, a drink, a bar, aprepared food contained in a can, a milk shake drink, a juice, a dairyfood product, or any other functional food composition. The foodcomposition may also comprise any form of a supplement such as a pill,soft gel, gummy figurine, wafer, or the like.

A food composition ingested by the mammal in addition to the proteinbuilding composition may also be rich in L-carnitine, leucine, and/orcreatine. The protein building composition of the present disclosure,for instance, is intended to provide additional L-carnitine, leucine,and/or creatine in addition to the normal amounts contained in astandard diet and/or the amounts produced by the body.

The mammal treated in accordance with the present disclosure cancomprise any suitable mammal. For instance, the mammal may be human orcanine. The protein building composition can be fed to a mammal of anyage such as from parturition through the adult life in the mammal. Invarious embodiments the mammal may be a human, dog, a cat, a horse, apig, a sheep, or a cow. In many embodiments, the mammal can be in earlyto late adulthood. For instance, the active mammal may have an age thatis at least 10%, such as least 15%, such as least 20%, such as least25%, such as least 30%, such as least 35%, such as least 40%, such asleast 45%, such as least 50%, such as least 55%, such as least 60%, suchas least 65%, such as least 70%, such as least 75%, such as least 85%,such as least 90%, such as least 95% of its expected life span. Themammal may have an age such that it is less than about 95%, such as lessthan about 90%, such as less than about 85%, such as less than about80%, such as less than about 75%, such as less than about 70%, such asless than about 65%, such as less than about 60%, such as less thanabout 55%, such as less than about 50%, such as less than about 45%,such as less than about 40%, such as less than about 35%, such as lessthan about 30%, such as less than about 25%, such as less than about20%, such as less than about 15%, such as less than about 10% of itsexpected life span. A determination of life span may be based onactuarial tables, calculations, or the like.

The protein building composition may be administered to the mammalaccording to the present disclosure regardless of the frequency,intensity, or type of physical activity performed by the mammal. Themammal may participate in physical activities with various MET values.In one embodiment, the mammal may regularly participate in light tointense physical activity. Light physical activity may have a MET offrom about 3 MET to about 6 MET. Moderate physical activity may have aMET of from about 6 MET to about 10 MET. Intense physical activity mayhave a MET of about 10 MET or greater. In another embodiment, the mammalmay infrequently participate in physical activity. In yet anotherembodiment, the mammal may lead a sedentary lifestyle, wherein themammal may rarely or never participate in physical activity. In asedentary lifestyle, a mammal may participate mainly or exclusively insedentary activities.

The protein building composition may be administered to the mammalbefore, during, or after a period of physical activity. Alternately, thecomposition may be administered to the mammal before, during, or after aperiod of sedentary activity. For instance, the composition may beadministered to the mammal during an extended period of bed rest orother extended period of inactivity.

The protein building composition is administered in an amount sufficientto increase muscle protein synthesis, increase functional strength, orincrease both muscle protein synthesis and functional strength withoutrequiring the mammal to participate in physical activity.

Muscle protein synthesis, in one embodiment, can be determined bymonitoring the biomarkers, mTOR expression and phosphorylation and itsrelated upstream and downstream proteins in the pathway, in skeletalmuscle. Specifically, mTOR expression can be determined and recordedbefore and after a period of activity. For a mammal treated inaccordance with the present disclosure, mTOR expression before and aftera period of time may vary by more than 10%, such as by more than 20%,such as by more than 40%, such as by more than 60%, such as by more than80%, such as by more than 100%, such as by more than 150%, such as bymore than 200%.

In one embodiment, the mammals treated in accordance with the presentdisclosure may have total mTOR values after a period of activity thatare at least 10%, such as at least 20%, such as at least 30%, such as atleast 40%, such as at least 50%, such as at least 60%, such as at least100% greater than the same mammal that is not administered the proteinbuilding composition.

Protein synthesis can be monitored by androgens, androgen receptors,insulin, IGF-1, IGF-1 receptors and any known stimulator of proteinsynthesis.

In addition to increased muscle protein synthesis, the protein buildingcomposition can also increase functional strength. In particular, theprotein building composition can increase lean muscle mass and upper andlower body strength. Functional strength can be measured by a compositeendpoint of strength and muscle measures. The composite endpoint may bethe product of the values for muscle mass (kg), upper extremity strengthby dynamometer (kg), lower extremity strength by dynamometer (kg), and6-minute walk test (meters). Comparative measurements can be taken priorto and after a period of activity.

In one embodiment, mammals treated in accordance with the presentdisclosure may have composite endpoints after a certain period of timethat are at least 10%, such as at least 25%, such as at least 50%, suchas at least 75%, such as at least 100% more than the same mammal that isnot administered the L-carnitine supplement.

The protein building composition of the present disclosure may alsoreduce inflammation. Inflammation, in one embodiment, can be determinedby monitoring the biomarker, TNF-α, in skeletal muscle. TNF-α is aninflammatory marker involved in protein degradation. Reduction in TNF-αvalues may indicate a reduction in protein degradation and musclewasting. TNF-α values can be determined and recorded before and after aperiod of activity. For a mammal treated in accordance with the presentdisclosure, TNF-α values before and after a period of time may vary bymore than 0.5%, such as by more than 1%, such as by more than 5%, suchas by more than 10%, such as by more than 20%, such as by more than 40%,such as by more than 60%, such as by more than 80%, such as by more than100%, such as by more than 150%, such as by more than 200%.

In one embodiment, the mammals treated in accordance with the presentdisclosure may have total TNF-α values after a period of activity thatare at least 0.5%, such as at least 1%, such as at least 5%, such as atleast 10%, such as at least 20%, such as at least 30%, such as at least40%, such as at least 50%, such as at least 60%, such as at least 100%less than the same mammal that is not administered the protein buildingcomposition.

Reduced inflammation can lead to decreased protein degradation.

In one embodiment, the compositions of the present disclosure maycontain other amino acids, including but not limited to alanine,arginine, asparagine, aspartate, cysteine, glutamic acid, glutamine,glycine, proline, serine, tyrosine, histidine, isoleucine, lysine,methionine, phenylalanine, threonine, tryptophan, valine, and anycombinations thereof.

The protein building composition of the present disclosure may furthercomprise one or more excipients. Exemplary but non-limiting excipientsinclude antiadherents, such as magnesium stearate; binders, such assaccharides, sugar alcohols, gelatin, and synthetic polymers; coatings,such as cellulose ether hydroxypropyl methylcellulose (HPMC), shellac,corn protein zein, gelatin, fatty acids, and waxes; coloring agents,such as titanium oxide and azo dyes; disintegrants, such as modifiedstarch sodium starch glycolate and crosslinked polymers includingpolyvinylpyrrolidone and sodium carboxymethyl cellulose; fillers, suchas maltodextrin; flavoring agents, such as mint, liquorice, anise,vanilla, and fruit flavors including peach, banana, grape, strawberry,blueberry, raspberry, and mixed berry; glidants, such as fumed silica,talc, and magnesium carbonate; lubricants, such as talc, silica, andfats including vegetable stearin, magnesium stearate, and stearic acid;preservatives, such as antioxidants, vitamins, retinyl palmitate,selenium, the amino acids cysteine and methionine, citric acid, sodiumcitrate, and parabens; sorbents; sweeteners, such as sucrose andsucralose; and vehicles, such as petrolatum and mineral oil.

In one embodiment, the protein building composition of the presentdisclosure may be combined with various additives and components thatcan improve one or more properties of the composition. For example, inone embodiment, the additive composition may be combined with astabilizer package that may serve to stabilize at least one property ofthe composition. In one particular embodiment, for instance, astabilizer package may be added to the composition in an amountsufficient to reduce the hydroscopic properties of the compositionand/or prevent the composition from absorbing moisture. A stabilizerpackage may also be combined with the protein building composition inorder to improve the handling properties of the composition. Forinstance, the stabilizer package may allow the composition to havebetter flow properties, especially when in granular form.

In one embodiment, the protein building composition may be combined witha polymer binder in conjunction with a stabilizer package. In addition,a coating material may also be applied to the composition after thecomposition has been combined with the polymer binder and the stabilizerpackage. The coating material, for instance, may contain at least onefat. In accordance with the present disclosure, the above components canbe added to any suitable pharmaceutical composition in addition to theprotein building composition of the present disclosure. For instance,the above components may be added to any pharmaceutical compositioncontaining a carnitine or an amino acid.

The polymer binder and the stabilizer package may be combined with theprotein budding composition in a manner that homogeneously incorporatesthe stabilizer package into the product. In one embodiment, forinstance, the protein building composition of the present disclosure isfirst combined with a polymer binder, such as through a spray dryprocess, and then combined with the stabilizer package. The polymerbinder may comprise any suitable pharmaceutically acceptable polymer,such as film-forming polymers and/or polysaccharides. Particularexamples of polymer binders that may be used in accordance with thepresent disclosure include starch, maltodextrin, gum arabic,arabinogalactan, gelatin, and mixtures thereof. In one embodiment, thepolymer binder is added to the pharmaceutical composition in an amountof at least about 5% by weight, such as at least about 8% by weight,such as at least about 10% by weight, such as at least about 15% byweight. One or more polymer binders are present in the composition in anamount less than about 50% by weight, such as in an amount less thanabout 45% by weight, such as in an amount less than about 40% by weight,such as in an amount less than about 35% by weight, such as in an amountless than about 30% by weight.

In one embodiment, the polymer binder may comprise a starch, such as amodified starch. The starch, for instance, may be derived from corn orwaxy maize. In one embodiment, the starch may comprise HI-CAP100 starchsold by National Starch and Chemical Company.

In an alternative embodiment, the polymer binder may comprisearabinogalactan. Arabinogalactan is a soluble polysaccharide that notonly can serve as a polymer binder but may also provide other benefits.For instance, arabinogalactan may enhance the adaptive immune responsein some circumstances. Arabinogalactan is described, for instance, inU.S. Pat. No. 8,784,844, which is incorporated herein by reference.

In one embodiment, larch arabinogalactan may be used as the polymerbinder. Larch arabinogalactan is a highly branched polysaccharide thatis composed of galactose units and arabinose units in the approximateratio of 6:1. Larch arabinogalactan is extracted from large trees. Thepolysaccharide has a galactan backbone with side chains of galactose andarabinose. Arabinogalactan is commercially available from Lonza Ltd.

Once the polymer binder is combined with the protein buildingcomposition such as through a spray dry process, the resulting mixturecan then be combined with a stabilizer package. In one embodiment, thestabilizer package comprises oxide particles in combination with a saltof a carboxylic acid. In one particular embodiment, the stabilizerpackage may comprise a dry product, such as a powder or granular productthat is combined with the protein building composition and polymerbinder. The combination of oxide particles and a salt of a carboxylicacid have been found to provide numerous advantages and benefits whencombined with the protein building composition. For instance, thestabilizer package has been found to stabilize the protein buildingcomposition and make the composition less hydroscopic. The compositionis also easier to handle and, when in granular form, produces afree-flowing product.

The oxide particles that may be added to the pharmaceutical compositionmay comprise silica. For instance, the oxide particles may compriseprecipitated silica particles. The silica particles may have a particlesize (d50, laser detraction following ISO Test 13320) of less than about55 microns, such as less than about 40 microns, such as less than about30 microns, such as less than about 25 microns, such as less than about20 microns, such as less than about 15 microns, such as less than about12 microns, such as less than about 10 microns, such as less than about8 microns, such as less than about 6 microns, such as less than about 4microns, such as less than about 2 microns, such as less than about 1micron. The particle size is typically greater than about 0.5 microns,such as greater than about 1 micron. The particles may have a specificsurface area (ISO Test 9277) of greater than about 120 m²/g, such asgreater than about 130 m²/g, such as greater than about 150 m²/g, suchas greater than about 170 m²/g, such as greater than about 200 m²/g,such as greater than about 220 m²/g. The specific surface area isgenerally less than about 500 m²/g. The oxide particles, such as thesilica particles, can be present in the pharmaceutical composition in anamount greater than about 0.01% by weight, such as in an amount greaterthan about 0.05% by weight, such as in an amount greater than about 0.1%by weight. The oxide particles are generally present in an amount lessthan 5% by weight, such as in an amount less than about 2% by weight,such as in an amount less than about 1.5% by weight, such as in anamount less than 0.5% by weight.

In addition to the oxide particles, the stabilizer package may alsoinclude a salt of a carboxylic acid. The salt of a carboxylic acid maycomprise a salt of a fatty acid. The fatty acid, for instance, may havea carbon chain length of from about 6 carbon atoms to about 40 carbonatoms, such as from about 12 carbon atoms to about 28 carbon atoms. Inone embodiment, the salt of the carboxylic acid may comprise a stearatesalt. The stearate salts that may be used include calcium stearate,sodium stearate, magnesium stearate, mixtures thereof, and the like. Inone embodiment, the salts of the carboxylic acid may include bothhydrophilic groups and hydrophobic groups. The salt of the carboxylicacid may be present in the pharmaceutical composition in an amountgreater than about 0.5% by weight, such as in an amount greater thanabout 1% by weight, such as in an amount greater than about 1.5% byweight. The salt of the carboxylic acid is generally present in anamount less than about 5% by weight, such as in an amount less thanabout 4% by weight, such as in an amount less than about 3% by weight.

In addition to the polymer binder and the stabilizer package, thecomposition may include various other components and ingredients. In oneembodiment, for instance, the composition may contain a citric acidester, such as a citric acid ester of a mono and/or diglyceride of afatty acid. The composition may also contain a lecithin, such as alecithin obtained from rapeseed, sunflower, and the Ike. The abovecomponents can be present in the composition in relatively minoramounts, such as less than about 2% by weight, such as less than about1.5% by weight, such as less than about 1% by weight. The abovecomponents are generally present in an amount greater than about 0.05%by weight, such as in an amount greater than about 0.1% by weight.

Once the above components are combined together to form thepharmaceutical composition, the composition can optionally be combinedwith a coating material. In one embodiment, for instance, thepharmaceutical composition may comprise a granular composition to whicha coating material is applied that contains a fat. The coating material,for instance, may comprise a hydrogenated oil, such as hydrogenated palmoil. In one particular embodiment, the coating material may comprisehydrogenated palm oil combined with palm stearine. In one embodiment,the hydrogenated oil may be present in the pharmaceutical composition inan amount from about 5% to about 35% by weight. The palm stearine, onthe other hand, may be present in the pharmaceutical composition in anamount from about 2% to about 10% by weight. When present together, theweight ratio between the hydrogenated palm oil and the palm stearine maybe from about 10:1 to about 1:1, such as from about 6:1 to about 2:1. Inone embodiment, the hydrogenated palm oil and the palm stearine arepresent at a weight ratio of about 4:1.

The present disclosure may be better understood with reference to thefollowing examples.

Example No. 1

A randomized, double-blind, placebo-controlled study was conducted toevaluate the effects of two protein building compositions formulatedaccording to the present disclosure on muscle protein synthesis,functional strength, lean body mass, and overall quality of life inhealthy older adults.

The study was conducted with a sample size of 42 healthy older adults,with 14 subjects randomized to each study arm in a double-blind mannerat a ratio of 1:1:1. Each subject was selected for compliance with theinclusion and exclusion criteria shown in Tables 1 and 2.

Table 1. Inclusion Criteria For Test Subjects

Healthy male or female adults, aged 55 to 70 yearsBMI of 21 kg/m² to 33 kg/m²Subjects in good physical condition such that they can perform exercisetesting safely, as determined by the Qualified Investigator based onmedical history, physical examination, electrocardiogram and laboratoryresultsSubjects who are sedentary and not currently engaging in any regularexercise.Subjects who agree to maintain their current level of activity andcurrent dietary habits throughout the trial period.Subjects who have given voluntary, written, informed consent toparticipate in the study.

TABLE 2 Exclusion Criteria For Test Subjects Subjects who are smokers orhave been a smoker within the past 1 year from screening. Subjects whoare pregnant or breastfeeding Subjects who have experienced weight lossor gain of greater than 4.5 kg (approximately 10 lbs) within 3 months ofrandomization Subjects diagnosed with active heart disease Subjects withuncontrolled hypertension 140 mmHg) Subjects with renal or hepaticimpairment or disease Subjects with any major diseases of thegastrointestinal, pulmonary or endocrine systems Subjects with a historyof seizures Subjects with Type I and Type II Diabetes Subjects withactive cancer (excluding basal cell carcinoma) Subjects withneurological or significant psychiatric illnesses, including Parkinson'sdisease and bi-polar disorder Subjects with unstable thyroid diseaseSubjects who are immuno-compromised (HIV positive, on anti-rejectionmedication, rheumatoid arthritis) Subjects with metal fixation plates orscrews from a previous surgery Subjects who are taking oralanticoagulants (blood thinners) such as warfarin (Coumadin) orDabigatran (Pradaxa) or antiplatelet agents such as Clopidogrel (Plavix)Subjects who are regularly taking NSAID medications such as aspirin,must stop at least one week prior to the micro-needle muscle biopsyprocedures. Subjects with a known allergy to anesthetic Subjects whocurrently experience any medical condition that interferes with theability to undergo physical strength testing during the study Subjectscurrently taking NHPs must have been using their current dosing regimenfor at least one month prior to baseline and must maintain their currentdosing regimen throughout the trial and must not begin taking any newNHPs throughout the trial. If the subject wishes to stop taking the NHPprior to beginning the trial, they must do so at least 1 week prior torandomization. Subjects who use illicit drugs or have a history ofalcohol or drug abuse within the past 6 months Subjects who currentlyconsume greater than 2 standard alcoholic drinks per day. Subjects whohave participated in a clinical research trial within 30 days prior torandomization. Subjects with an allergy or sensitivity to theinvestigational product ingredient. Subjects who are cognitivelyimpaired and/or who are unable to give informed consent Subjects whohave abnormal laboratory results or any other medical or psychologicalcondition which, in the opinion of the Qualified Investigator, mayadversely affect the subject's ability to complete the study or itsmeasures or which may pose significant risk to the subject.

The compositions of the three nutritional supplements are shown in Table3. Subjects received one of the three supplements based on whether theywere in the Sample 1, Sample 2, or the placebo group. The Sample 1 groupreceived 4 grams of the designated composition per day. The Sample 2group received 8 grams of the designated composition per day. Theplacebo group received 3 grams of the designated composition per day.Subjects were instructed to empty the contents of the sachet of thedesignated composition into one bottle of orange juice and shakevigorously for 1 min, then drink the entire bottle. Each sachet forSample 1 and Sample 2 contained 2.2 g of L-carnitine tartrate. Subjectswere instructed to start taking the product the day after randomization(Day 1). The subjects continued to take the product for an 8 weekperiod.

Subjects were asked to maintain their current physical activity levelsand dietary habits throughout the trial.

TABLE 3 L-Carnitine and Placebo Supplement Compositions ActiveIngredients Other Ingredients Sample 1 L-carnitine (1500 mg)Maltodextrin Sucrose Colloidal Silicon Dioxide Mixed Berry FlavorSucralose Sample 2 L-carnitine (1500 mg) Sucrose L-Leucine (2000 mg)Colloidal Silicon Dioxide Creatine monohydrate Mixed Berry Flavor (3000mg) Vitamin D3 (4001 U) Sucralose Placebo Tartaric Acid MaltodextrinSucrose Colloidal Silicon Dioxide Mixed Berry Flavor Sucralose

Assessments were conducted at Day 0, Day 29±3, and Day 57±3 of thestudy. On Day 29, subjects were contacted via phone to monitor adverseevents and changes in concomitant medications as well as to fostercompliance with the study dosing regimen.

Data was analyzed for the per protocol population, which consisted ofall participants who consumed at least 80% of treatment or placebodoses, did not have any major protocol violations and completed allstudy visits and procedures connected with measurement of the primaryvariable. The per-protocol population contained 39 subjects, as 3subjects were excluded from the original 42 subject population forprotocol violations. For statistical analysis, within-group changes over8 weeks of supplementation, and pre-to-post-strength testing changes,were tested for significance by the paired Student t test or thenon-parametric Wilcoxon Signed-ranks test. Between-group differences in8-week changes were tested for significance by the unpaired Student ttest or the non-parametric Mann-Whitney U test. In the following tables,treatment groups with differing letter superscripts are significantlydifferent. Probability values P≤0.05 are statistically significant.

Total lean mass was measured for each participant on Day 0 and Day 57±3via dual-energy X-ray absorptiometry (DXA). As shown in FIG. 1 and Table4, total lean mass was significantly increased (+1.01 kg, P=0.013) inthe Sample 2 group, but not in Sample 1 (−0.03 kg, P=0.915) or theplacebo (0.0 kg, P=0.986) groups.

As revealed in FIG. 2, arm lean mass significantly differed betweenSample 1 and 2 (P=0.012) by the end of 8 weeks, with Sample 1 showing aslight reduction (−0.137 kg) in arm mass and Sample 2 demonstrating aminor increase (+0.135 kg) in arm mass.

FIG. 3 demonstrated that participants supplemented with Sample 2 alsohad enhanced leg muscle mass (+0.35 kg, P=0.005), which was notreflected in the placebo group (−0.03 kg, P=0.811). Subjects on Sample 1did exhibit a small, but non-significant increase in leg lean mass(+0.17 kg, P=0.256) as well.

Illustrated in FIG. 4, total non-trunk lean mass increased significantlyin the Sample 2 group by the end of the study (+0.48 kg, P=0.006). Thischange was significantly greater than the placebo group (P<0.05), whichtended to lose total non-trunk lean mass (−0.10 kg, P=0.560). Sample 1group maintained their total non-trunk lean mass for the duration of thestudy (+0.03 kg, P=837).

TABLE 4 DXA Lean Masses at Baseline and at End of the Study for AllParticipants in the PP Population (N = 39). Placebo Sample 1 Sample 2Mean ± SD (n) Mean ± SD (n) Mean ± SD (n) Between Group P Values Median(Min − Median (Min − Median (Min − Placebo Placebo Sample 1 Max) Max)Max) vs vs vs Within Group P Within Group P Within Group P Over- SampleSample Sample 2 Value^(‡) Value^(‡) Value^(‡) all ^(Δ) 1^(δ) 2^(δ) ^(δ)Total Lean Mass (kg) Baseline 43.7 ± 9.1 (14) 44.8 ± 8.8 (11) 45.2 ± 8.6(14) — — — — (Week 0) 40.8 (30.4 − 60.1) 40.5 (33 − 60.1) 42.2 (34.9 −65.2) Visit 3 43.7 ± 9.2 (14) 45.0 ± 8.9 (11) 46.2 ± 9.7 (14) — — — —(Week 8) 40.8 (29.7 − 60.6) 40.9 (33.5 − 61.6) 43 (35 − 68.6) Change−0.00 ± 1.04 (14) 0.14 ± 1.22 (11) 1.01 ± 1.30 (14) 0.074 0.978 0.0880.169 from 0.23 (−2.11 − 1.36) 0.48 (−1.61 − 1.6) 0.98 (−1.37 − 3.41)Baseline p = 0.986 p = 0.716 p = 0.013 to Week 8 Arm Lean Mass (kg)Baseline 4.91 ± 1.47 (14) 5.11 ± 1.72 (11) 4.93 ± 1.43 (14) — — — —(Week 0) 4.8 (3.29 − 7.69) 4.11 (3.21 − 7.42) 4.4 (3.5 − 7.9) Visit 34.84 ± 1.40 (14) 4.99 ± 1.68 (11) 5.07 ± 1.52 (14) — — — — (Week 8) 4.37(3.06 − 7.22) 4.09 (3.12 − 7.3) 4.66 (3.39 − 7.98) Change −0.071 ± 0.276(14) −0.123 ± 0.088 (11) 0.135 ± 0.252 (14) 0.019 0.863 0.061 0.026 from0.015 (−0.55 − 0.42) −0.12 (−0.3 − 0.02) 0.15 (−0.45 − 0.48) Baseline p= 0.351 p < 0.001 p = 0.067 to Week 8 Leg Lean Mass (kg) Baseline 14.59± 3.13 (14) 14.30 ± 2.88 (11) 14.41 ± 3.06 (14) — — — — (Week 0) 13.98(10.92 − 20.34) 13.07 (10.91 − 19.02) 13.87 (11.29 − 20.98) Visit 3 14.6± 3.2 (14) 14.6 ± 3.0 (11) 14.8 + 3.2 (14) — — — — (Week 8) 13.6 (10.2 −20.1) 13.1 (11.7 − 20.1) 14.1 (11.1 − 21.6) Change −0.03 ± 0.46 (14)0.29 ± 0.50 (11) 0.35 ± 0.39 (14) 0.069 0.185 0.076 0.947 from −0.05(−0.73 − 0.7) 0.16 (−0.49 − 1.05) 0.38 (−0.35 − 0.99) Baseline p = 0.811p = 0.086 p = 0.005 to Week 8 Total Non-Trunk Lean Mass (kg) Baseline19.5 ± 4.5 (14) 19.4 ± 4.5 (11) 19.3 ± 4.4 (14) — — — — (Week 0) 18.4(14.2 − 27.7) 16.8 (14.2 − 26.4) 17.9 (14.8 − 28.9) Visit 3 19.4 ± 4.6(14) 19.6 ± 4.6 (11) 19.8 ± 4.7 (14) — — — — (Week 8) 17.9 (13.3 − 27)16.8 (14.8 − 27.4) 18.4 (14.5 − 29.5) Change −0.10 ± 0.64 (14) 0.16 ±0.51 (11) 0.48 ± 0.56 (14) 0.037 0.499 0.029 0.360 from 0.02 (−1.2 −1.12) 0.14 (−0.67 − 0.93) 0.54 (−0.48 − 1.22) Baseline p = 0.560 p =0.315 p = 0.006 to Week 8 Trunk Lean Mass (kg) Baseline 23.2 ± 10.8 (14)23.9 ± 6.4 (11) 27.6 ± 11.0 (14) — — — — (Week 0) 20.1 (13.9 − 58) 23.2(16.4 − 39.3) 23.6 (17.5 − 57.4) Visit 3 22.4 ± 6.2 (14) 22.1 ± 4.0 (11)23.0 ± 4.1 (14) — — — — (Week 8) 20.2 (14 − 37.9) 22.2 (16.1 − 29.9)22.5 (17.7 − 31.9) Change −0.8 ± 10.1 (14) −1.7 ± 5.1 (11) −4.6 ± 9.9(14)  0.943*  0.969*  0.942*  0.997* from 0.1 (−30.9 − 19.3) −0.4 (−17.1− 0.6) −0.2 (−28.4 − 1.7) Baseline p = 0.973* p = 0.271* p = 0.101* toWeek 8 N, number; SD, standard deviation; Min, minimum; Max, maximum;kg, kilogram ^(Δ) Between-group comparisons were made using ANCOVA.^(δ)Pairwise between-group comparisons were made using the Tukeyprocedure. Within-group comparisons were made using the paired Studentt-test. *Logarithmic transformation was required to achieve normality.Treatment groups with differing letter superscripts are significantlydifferent. Probability values P ≤ 0.05 are statistically significant.

Lower leg strength was assessed by average leg dynamometry for eachparticipant on Day 0 and Day 57±3. As shown in FIG. 5 and Table 5, lowerleg strength was significantly increased in the Sample 2 group (+1.0 kg,P=0.029). This increase in leg dynamometry corresponds with the rise inleg muscle mass (DXA scan) and was significantly different from theplacebo group (P=0.012), which exhibited reduced average leg strength(−2.8 kg, P=0.061) by the end of study. Sample 1 significantly differedfrom the placebo group (P=0.023) and maintained their average legstrength throughout the study duration (−0.4 kg, P=0.764). Upper bodystrength markers, including average left and right hand, as well asaverage arm strength did not significantly change for any treatmentgroup.

TABLE 5 Dynamometry Results at Baseline and at End of the Study for AllParticipants in the PP Population (N = 39). Placebo Sample 1 Sample 2Mean ± SD (n) Mean ± SD (n) Mean ± SD (n) Between Group P Values Median(Min − Median (Min − Median (Min − Placebo Placebo Sample 1 Max) Max)Max) vs vs Vs Within Group P Within Group P Within Group P Over- SampleSample Sample Value Value Value all ^(Δ) 1^(δ) 2^(δ) 2^(δ) Average RightHand (kg) Baseline 28.3 ± 10.3 (14) 26.5 ± 10.3 (11) 24.0 ± 9.9 (14) — —— — (Week 0) 27.5 (4.5 − 45.6) 26.3 (11.2 − 45.9) 26 (7.6 − 40.8) Visit3 28.6 ± 11.6 (14) 24.9 ± 10.7 (11) 25.6 ± 9.4 (14) — — — — (Week 8)25.2 (3.9 − 49.8) 23.3 (10.3 − 42.3) 25.7 (10 − 41.1) Change 0.3 ± 4.9(14) −1.5 ± 4.2 (11) 1.7 ± 6.3 (14) 0.383 0.619 0.884 0.358 from −0.7(−4.8 − 14.2) 0.3 (−11.6 − 4.2) 0.3 (−6.8 − 19.5) Baseline p = 0.819 p =0.257 p = 0.345 to Week 8 Average Left Hand (kg) Baseline 27.1 ± 11.2(14) 26.2 ± 12.1 (11) 23.8 ± 10.6 (14) — — — — (Week 0) 26.8 (4.7 −48.2) 24.8 (9.4 − 52.1) 25.3 (8.3 − 41.5) Visit 3 27.3 ± 10.2 (14) 25.8± 12.1 (11) 26.2 ± 9.6 (14) — — — — (Week 8) 26.2 (5.1 − 42) 26.1 (9.8 −50.9) 25.3 (10.1 − 40.8) Change 0.2 ± 4.3 (14) −0.3 ± 4.1 (11) 2.3 ± 4.9(14) 0.363 0.921 0.552 0.366 from 1.1 (−7.6 − 7.7) −0.6 (−10.6 − 4.1)1.9 (−3.3 − 16.2) Baseline p = 0.869 p = 0.791 p = 0.095 to Week 8Average Arm (kg) Baseline 27.7 ± 10.7 (14) 26.3 ± 11.2 (11) 23.9 ± 10.1(14) — — — — (Week 0) 27.1 (4.6 − 46.9) 25.7 (10.3 − 49) 26.1 (7.9 −41.1) Visit 3 27.9 ± 10.8 (14) 25.4 ± 11.3 (11) 25.9 + 9.3 (14) — — — —(Week 8) 25.3 (4.5 − 45.9) 24.1 (10 − 46.6) 25.6 (10 − 39.9) Change 0.23± 4.2 (14) −0.9 ± 4.0 (11) 2.0 ± 5.3 (14) 0.354 0.744 0.725 0.322 from 0(−5.5 − 10) −0.1 (−11.1 − 4.2) 0.9 (−3.8 − 17.8) Baseline p = 0.828 p =0.455 p = 0.186 to Week 8 Average Leg (kg) Baseline 13.5 ± 6.4 (14) 15.7± 5.1 (11) 12.3 ± 7.2 (14) — — — — (Week 0) 12.4 (4.5 − 24.5) 16.8 (6.9− 23.6) 10.5 (4.5 − 24.6) Visit 3 10.7 ± 3.6 (14) 15.7 ± 5.1 (11) 13.4 ±6.2 (14) — — — — (Week 8) 9.9 (6.5 − 19.3) 15.1 (9.4 − 26.6) 13.1 (6 −23.3) Change −2.8 ± 4.8 (14)^(a) 0.0 ± 4.5 (11)^(b) 1.0 ± 2.0 (14)^(b) 0.006^(†)  0.016^(†)  0.014^(†)  0.988^(†) from −2 (−14.8 − 2.4) 1.1(−8.6 − 5.2) 1.2 (−3.8 − 4.5) Baseline p = 0.061^(†) p = 0.952^(†) p =0.029^(†) to Week 8 N, number; SD, standard deviation; Min, minimum;Max, maximum; kg, kilogram ^(Δ) Between-group comparisons were madeusing ANCOVA. ^(δ)Pairwise between-group comparisons were made using theTukey procedure. Within-group comparisons were made using the pairedStudent t-test. ^(†)Square root transformation required to achievenormality. Treatment groups with differing letter superscripts aresignificantly different. Probability values P ≤ 0.05 are statisticallysignificant.

The participants completed the six minute walk test on Day 0 and Day57±3. Table 6 shows that participants taking placebo had a significantreduction in their out of breath score after walking at the end of studyrelative to baseline (−0.43, P<0.05). There were no notable changes inthe total meters walked, out of breath score before walking, and fatiguescore before/after walking in all treatment groups.

TABLE 6 Six Minute Walk Test at Baseline and at End of the Study for AllParticipants in the PP Population (N = 39). Placebo Sample 1 Sample 2Mean ± SD (n) Mean ± SD (n) Mean ± SD (n) Between Group P Values Median(Min − Median (Min − Median (Min − Placebo Placebo Sample 1 Max) Max)Max) vs vs Vs Within Group P Within Group P Within Group P Over- SampleSample Sample Value Value Value all ^(Δ) 1^(δ) 2^(δ) 2^(δ) Meters Walkedin Six Minutes (m) Baseline 526 ± 80 (14) 458 ± 127 (11) 432 ± 109 (14)— — — — (Week 0) 542 (336 − 633) 442 (239 − 695) 460 (220 − 540) Visit 3530 ± 100 (14) 444 ± 119 (11) 462 ± 113 (14) — — — — (Week 8) 560 (220 −627) 497 (220 − 560) 490 (238 − 600) Change 3 ± 69 (14) −14 ± 107 (11)30 ± 70 (14) 0.310^(†) 0.326^(†) 0.961^(†) 0.449^(†) from −3 (−116 −140) −8 (−222 − 135) 27 (−120 − 138) Baseline p = 0.704^(†) p =0.617^(†) p = 0.126^(†) to Week 8 Out of Breath Score Before WalkingBaseline 0.00 ± 0.00 (14) 0.09 ± 0.30 (11) 0.21 ± 0.80 (14) — — — —(Week 0) 0 (0 − 0) 0 (0 − 1) 0 (0 − 3) Visit 3 0.000 ± 0.000 (14) 0.045± 0.151 (11) 0.036 ± 0.134 (14) — — — — (Week 8) 0 (0 − 0) 0 (0 − 0.5) 0(0 − 0.5) Change 0.00 ± 0.00 (14) −0.05 ± 0.15 (11) −0.18 ± 0.67 (14)0.555^(¤) 0.93^(¤)  0.94^(¤)  1.00^(¤)  from 0 (0 − 0) −0.12 (−0.3 −0.02) 0.15 (−0.45 − 0.48) Baseline p = 1.000^(‡) p = 1.000^(‡) p =1.000^(‡) to Week 8 Out of Breath Score After Walking Baseline 10.4 ±0.91 (14) 0.55 ± 0.79 (11) 1.07 ± 2.16 (14) — — — — (Week 0) 1 (0 − 3) 0(0 − 2) 0 (0 − 7) Visit 3 0.61 ± 0.56 (14) 0.41 ± 0.58 (11) 1.00 + 1.79(14) — — — — (Week 8) 0.5 (0 − 2) 0.5 (0 − 2) 0 (0 − 5) Change −0.43 ±0.70 (14) −0.14 ± 0.64 (11) −0.07 ± 0.87 (14) 0.347^(¤) 0.45^(¤) 0.46^(¤)  1.00^(¤)  from −0.5 (−2 − 0.5) 0 (−1.5 − 0.5) 0 (−2 − 2)Baseline p = 0.044^(‡) p = 0.590^(‡) p = 0.730^(‡) to Week 8 Change inOut of Breath Score After Walking Baseline 1.04 ± 0.91 (14) 0.45 ± 0.65(11) 0.86 ± 1.60 (14) — — — — (Week 0) 1 (0 − 3) 0 (0 − 2) 0 (0 − 5)Visit 3 0.61 ± 0.56 (14) 0.36 ± 0.45 (11) 0.96 ± 1.70 (14) — — — — (Week8) 0.5 (0 − 2) 0.5 (0 − 1.5) 0 (0 − 5) Change −0.43 ± 0.70 (14) −0.09 ±0.66 (11) 0.11 ± 0.68 (14) 0.155^(¤) 0.33^(¤)  0.20^(¤)  0.98^(¤)  from−0.5 (−2 − 0.5) 0 (−1.5 − 0.5) 0 (−1 − 2) Baseline p = 0.044^(‡) p =0.792^(‡) p = 0.792^(‡) to Week 8 Fatigue Score Before Walking Baseline0.54 ± 0.93 (14) 0.50 ± 1.02 (11) 0.21 ± 0.54 (14) — — — — (Week 0) 0 (0− 3) 0 (0 − 3) 0 (0 − 2) Visit 3 0.39 ± 0.92 (14) 0.18 ± 0.60 (11) 0.25± 0.80 (14) — — — — (Week 8) 0 (0 − 3) 0 (0 − 2) 0 (0 − 3) Change −0.14± 0.57 (14) −0.32 ± 1.27 (11) 0.04 ± 0.95 (14) 0.929^(¤) 0.98^(¤) 0.99^(¤)  0.94^(¤)  from 0 (−1 − 1) 0 (−3 − 2) 0 (−1.5 − 3) Baseline p =0.387^(‡) p = 0.461^(‡) p = 0.854^(‡) to Week 8 Fatigue Score AfterWalking Baseline 0.82 ± 0.91 (14) 0.82 ± 1.03 (11) 0.64 ± 1.36 (14) — —— — (Week 0) 0.5 (0 − 3) 0.5 (0 − 3) 0 (0 − 5) Visit 3 0.61 ± 0.86 (14)0.45 ± 0.88 (11) 0.93 ± 1.25 (14) — — — — (Week 8) 0.5 (0 − 3) 0 (0 − 3)0.25 (0 − 3) Change −0.21 ± 0.64 (14) −0.36 ± 1.38 (11) 0.29 ± 1.17 (14)0.254^(¤) 0.98^(¤)  0.38^(¤)  0.33^(¤)  from −0.25 (−1.5 − 1) 0 (−3 −2.5) 0 (−2 − 3) Baseline p = 0.266^(‡) p = 0.348^(‡) p = 0.394^(‡) toWeek 8 Change in Fatigue Score After Walking Baseline 0.29 ± 0.64 (14)0.32 ± 0.64 (11) 0.43 ± 1.33 (14) — — — — (Week 0) 0.25 (−1 − 2) 0 (−0.5− 2) 0 (0 − 5) Visit 3 0.21 ± 0.26 (14) 0.27 ± 0.34 (11) 0.68 ± 1.05(14) — — — — (Week 8) 0 (0 − 0.5) 0 (0 − 1) 0 (0 − 3) Change −0.07 ±0.68 (14) −0.05 ± 0.65 (11) 0.25 ± 1.03 (14) 0.437^(¤) 0.92^(¤) 0.47^(¤)  0.75^(¤)  from 0 (−1.5 − 1.5) 0 (−1.5 − 1) 0 (−2 − 2.5)Baseline p = 0.660^(‡) p = 1.00^(‡) p = 0.34^(‡) to Week 8 N, number;SD, standard deviation; Min, minimum; Max, maximum ^(Δ) Between-groupcomparisons were made using ANCOVA. ^(δ)Pairwise between-groupcomparisons were made using the Tukey procedure. Within-groupcomparisons were made using the paired Student t-test. ^(†)Square roottransformation required to achieve normality. ^(¤)Between-groupcomparisons were made using the Kruskal Wallis test. ^(‡)Within-groupcomparisons were made using the signed-rank test. Probability values P ≤0.05 are statistically significant.

Quality of life was determined by the participants' responses to theRAND SF-36 questionnaire on Day 0 and Day 57±3 shown in Table 7. TheSF-36 questionnaire was developed at RAND as part of the MedicalOutcomes Study. The Sample 1 group showed a trend towards increasingtheir energy/fatigue ratio (+6.1, P=0.073) by the end of the studyrelative to baseline. All other quality of life measures, includingphysical functioning, role functioning (physical or emotional),emotional well-being, social functioning, pain, and general health werenot significantly changed by any intervention.

TABLE 7 SF-36 Questionnaire Results at Baseline and at End of the Studyfor All Participants in the PP Population (N = 39). Placebo Sample 1Sample 2 Mean ± SD (n) Mean ± SD (n) Mean ± SD (n) P Value^(¤) Median(Min − Median (Min − Median (Min − Placebo Placebo Sample 1 Max) Max)Max) vs vs Vs Within Group Within Group Within Group Over- Sample SampleSample 2 P Value^(‡) P Value^(‡) P Value^(‡) all^(¤) 1^(δ) 2^(δ) ^(δ)Physical Functioning Baseline 88.6 ± 16.2 (14) 86.4 ± 15.2 (11) 81.1 ±19.0 (14) — — — — (Week 0) 97.5 (50 − 100) 95 (60 −100) 90 (35 − 100)Visit 3 88.6 ± 14.6 (14) 85.9 ± 12.0 (11) 80.7 ± 13.4 (14) — — — — (Week8) 97.5 (65 − 100) 85 (60 − 100) 82.5 (50 − 100) Change from 0.0 ± 8.3(14) −0.5 ± 7.2 (11) −0.4 ± 14.9 (14) 0.713 0.78  0.21  0.63  Baselineto 0 (−15 − 20) 0 (−15 − 15) −2.5 (−25 − 35) Week 8 p = 1.000 p = 0.890p = 0.720 Role Functioning/Physical Baseline 90.2 ± 19.7 (14) 97.7 ± 5.1(11) 90.2 ± 17.1 (14) — — — — (Week 0) 100 (50 − 100) 100 (87.5 − 100)100 (50 − 100) Visit 3 93.8 ± 12.7 (14) 95.5 ± 11.6 (11) 92.9 ± 16.0(14) — — — — (Week 8) 100 (62.5 − 100) 100 (62.5 − 100) 100 (50 − 100)Change from 3.6 ± 19.9 (14) −2.3 ± 7.5 (11) 2.7 ± 25.6 (14) 0.596 0.99 1.00  0.98  Baseline to 0 (−25 − 50) 0 (−25 − 0) 0 (−50 − 50) Week 8 p =0.588 p = 1.000 p = 0.833 Role Functioning/Emotional Baseline 96.4 ±13.4 (14) 100.0 ± 0.0 (11) 97.6 ± 6.1 (14) — — — — (Week 0) 100 (50 −100) 100 (100 − 100) 100 (83.3 − 100) Visit 3 100.0 ± 0.0 (14) 93.9 ±13.5 (11) 95.2 ± 12.1 (14) — — — — (Week 8) 100 (100 − 100) 100 (66.7 −100) 100 (66.7 − 100) Change from 3.6 ± 13.4 (14) −6.1 ± 13.5 (11) −2.4± 14.4 (14) 0.319 0.72  0.79  0.99  Baseline to 0 (0 − 50) 0 (−33.3 − 0)0 (−33.3 − 16.7) Week 8 p = 1.000 p = 0.346 p = 0.577 Energy/FatigueBaseline 68.9 ± 21.0 (14) 68.2 ± 17.4 (11) 58.2 ± 20.1 (14) — — — —(Week 0) 72.5 (25 − 90) 70 (35 − 90) 60 (10 − 80) Visit 3 68.6 ± 16.5(14) 77.3 ± 11.7 (11) 57.5 ± 19.3 (14) — — — — (Week 8) 65 (35 − 100) 80(60 − 100) 60 (20 − 85) Change from −0.4 ± 19.6 (14)^(a,b) 9.1 ± 10.0(11)^(b) −0.7 ± 15.8 (14)^(a) 0.135 0.419 0.341 0.027 Baseline to 0 (−40− 45) 10 (−5 − 25) 0 (−45 − 15) Week 8 p = 0.780 p = 0.025 p = 0.691Emotional Well-Being Baseline 84.9 ± 15.2 (14) 86.5 ± 7.4 (11) 76.3 ±14.2 (14) — — — — (Week 0) 88 (40 − 100) 88 (76 − 100) 82 (48 − 96)Visit 3 84.3 ± 12.8 (14) 87.3 ± 12.2 (11) 81.1 ± 10.2 (14) — — — — (Week8) 86 (52 − 100) 92 (60 − 100) 84 (60 − 92) Change from −0.6 ± 13.8 (14)0.7 ± 10.1 (11) 4.9 ± 12.8 (14) 0.432 0.70  0.54  0.17  Baseline to 0(−32 − 36) 4 (−24 − 12) 4 (−12 − 28) Week 8 p = 0.670 p = 0.509 p =0.261 Social Functioning Baseline 51.8 ± 4.5 (14) 50.0 ± 5.6 (11) 48.2 ±8.3 (14) — — — — (Week 0) 50 (50 − 62.5) 50 (37.5 − 62.5) 50 (25 − 62.5)Visit 3 50.0 ± 0.0 (14) 52.3 ± 9.4 (11) 56.2 ± 19.5 (14) — — — — (Week8) 50 (50 − 50) 50 (37.5 − 75) 50 (25 − 100) Change from −1.8 ± 4.5 (14)2.3 ± 10.9 (11) 8.0 ± 20.0 (14) 0.368 0.93  0.78  0.96  Baseline to 0(−12.5 − 0) 0 (−12.5 − 25) 0 (−12.5 − 50) Week 8 p = 0.346 p = 0.572 p =0.202 Pain Baseline 84.1 + 18.3 (14) 83.4 ± 12.3 (11) 77.7 ± 15.1 (14) —— — — (Week 0) 90 (45 − 100) 80 (57.5 − 100) 78.8 (42.5 − 100) Visit 384.8 ± 14.6 (14) 83.0 ± 15.4 (11) 78.0 ± 20.6 (14) (Week 8) 90(6.75−100) 90 (57.5 −100) 85 (35 − 100) Change from 0.7 ± 11.0 (14) −0.5 ±13.0 (11) 0.4 ± 23.3 (14) 0.900 0.90  0.66  0.92  Baseline to 0 (−12.5 −22.5) 0 (−22.5 − 12.5) −5 (−32.5 − 57.5) Week 8 p = 1.000 p = 1.000 p =0.944 General Health Baseline 85.4 ± 14.3 (14) 85.9 ± 14.3 (11) 75.7 ±16.9 (14) — — — — (Week 0) 90 (55 − 100) 90 (50 − 100) 82.5 (40 − 95)Visit 3 83.2 ± 12.3 (14) 86.0 ± 8.2 (11) 77.5 ± 13.7 (14) — — — — (Week8) 80 (60 − 100) 85 (70 − 100) 80 (55 − 100) Change from −2.1 ± 11.4(14) 0.1 ± 12.2 (11) 1.8 ± 11.9 (14) 0.930 0.78  0.59  0.25  Baseline to0 (−30 − 10) 0 (−15 − 31.2) −2.5 (−15 − 30) Week 8 p = 0.892 p = 0.733 p= 0.662 N, number; SD, standard deviation; Min, minimum; Max, maximum^(¤)Between-group comparisons were made using the Kruskal Wallis test.^(δ)Pairwise between-group comparisons were made using the Tukeyprocedure. ^(‡)Within-group comparisons were made using the pairedStudent t-test. Probability values P ≤ 0.05 are statisticallysignificant.

For mRNA analysis, muscle biopsies were taken from participants (n=38)at Day 0 and at Day 57±3 to evaluate the effect of Sample 1 and 2 orplacebo on mRNA expression levels within the Ubiquitin-proteasomepathway, growth factor/protein signaling, and pro-inflammatory signalingpathways. mRNA was isolated from muscle tissue collected at baseline andend-of-study. From the mRNA, cDNA was synthesis and analyzed usingRT-PCR techniques. Anabolic/protein synthesis signaling genes (androgenreceptor, insulin receptor, IGF-1 & IGF-1 receptor), catabolic/proteindegradation signaling genes (atrogin-1 & MuFR1) and cytokines/cytokinereceptors (IL-6, IL-6 receptor, TNFα, TNFrSF1A, TNFrSF1B) genes wereassayed along with GAPDH as the endogenous control. The mRNA expressionfor the muscle biopsies collected suggests that the Samples affect boththe anabolic and catabolic pathways.

For the growth factor/protein signaling genes, Table 8 reveals thatthere were no significant between group differences observed; however,there was a trend for both Carnitine groups to have greatertranscription of these genes compared to placebo.

The subject number (N=38) for Table 8 is reflected by the processing andisolation of mRNA from subjects' muscle biopsies taken on Day 0 and Day57±3. Baseline and end of study muscle samples were required for thisanalysis (to see relative changes in the requested markers) and some ofthe subjects did not have enough skeletal muscle to yield the amount ofmRNA required to run the molecular analysis.

TABLE 8 Relative Changes in Quantity of Gene Transcription (mRNA) fromBaseline to End of the Study for All Participants in the PP Population(N = 38). Placebo Sample 1 Sample 2 Mean ± SD (n) Mean ± SD (n) Mean ±SD (n) P Value^(¤) Median (Min − Median (Min − Median (Min − PlaceboPlacebo Sample 1 Max) Max) Max) vs vs Vs Within Group Within GroupWithin Group Over- Sample Sample Sample 2 P Value P Value P Valueall^(¤) 1^(δ) 2^(δ) ^(δ) Androgen Receptor Relative 0.93 ± 0.43 (14)0.94 ± 0.37 (11) 1.10 ± 0.42 (13) 0.421* 0.529* 0.999* 0.459* Quantity0.82 (0.47 − 1.67) 1.01 (0.24 − 1.41) 1.08 (0.61 − 2.26) change in p =0.192* p = 0.335* p = 0.703* Gene Transcription (normalized to GAPDH)Baseline compared to End of Study Atrogin-1 Relative 0.98 ± 0.44 (14)1.51 ± 0.61 (11) 1.25 ± 0.46 (13) 0.030* 0.545* 0.025* 0.205* Quantity0.94 (0.38 − 1.68) 1.25 (0.97 − 2.68) 1.1 (0.57 − 2.17) change in p =0.375* p = 0.013* p = 0.142* Gene Transcription (normalized to GAPDH)Baseline compared to End of Study Insulin-like Growth Factor-1 Relative0.52 ± 0.28 (14) 0.74 ± 0.53 (11) 1.04 ± 0.72 (13) 0.122* 0.223* 0.990*0.143* Quantity 0.52 (0.14 − 1.23) 0.62 (0.04 − 1.52) 0.86 (0.19 − 2.82)change in p < 0.001* p = 0.070* p = 0.404* Gene Transcription(normalized to GAPDH) Baseline compared to End of Study Insulin-likeGrowth Factor-1 Receptor Relative 1.05 ± 0.65 (14) 1.37 ± 1.21 (11) 1.41± 0.86 (13) 0.647* 0.745* 0.996* 0.663* Quantity 0.99 (0.29 − 2.99) 1.21(0.11 − 4.51) 1.13 (0.3 − 3.03) change in p = 0.515* p = 0.817* p =0.422* Gene Transcription (normalized to GAPDH) Baseline compared to Endof Study IL-6 Relative 0.56 ± 0.37 (14) 1.64 ± 2.09 (11) 1.06 ± 0.69(13) 0.194* 0.870* 0.453* 0.183* Quantity 0.57 (0.05 − 1.3) 0.77 (0.03 −7.08) 0.8 (0.22 − 2.47) change in p = 0.005* p = 0.460* p = 0.473* GeneTranscription (normalized to GAPDH) Baseline compared to End of StudyIL-6 Receptor Relative 1.06 ± 1.07 (14) 1.29 ± 0.88 (11) 1.28 ± 0.68(13) 0.371* 0.949* 0.584* 0.368* Quantity 0.74 (0.27 − 4.28) 1.17 (0.31− 3.3) 1.2 (0.39 − 2.84) change in p = 0.249* p = 0.885* p = 0.455* GeneTranscription (normalized to GAPDH) Baseline compared to End of StudyInsulin Receptor Relative 0.97 ± 0.58 (14) 1.26 ± 1.36 (11) 1.18 ± 0.83(13) 0.813* 0.966* 0.933* 0.799* Quantity 1.02 (0.1 − 2.29) 0.91 (0.22 −4.96) 0.92 (0.09 − 3.49) change in p = 0.235* p = 0.556* p = 0.744* GeneTranscription (normalized to GAPDH) Baseline compared to End of StudyMuRF1 Relative 1.23 ± 0.76 (14) 2.00 ± 1.27 (11) 1.40 ± 1.10 (13) 0.092*0.178* 0.100* 0.957* Quantity 1.02 (0.48 − 3.24) 1.8 (0.9 − 5.44) 1.2(0.37 − 4.68) change in p = 0.679* p = 0.005* p = 0.503* GeneTranscription (normalized to GAPDH) Baseline compared to End of StudyTNF-α Relative 0.43 ± 0.37 (14) 0.76 ± 1.13 (11) 1.13 ± 1.17 (13) 0.054*0.188* 0.873* 0.053* Quantity 0.32 (0.02 − 1.28) 0.48 (0.02 − 3.99) 0.57(0.28 − 4.44) change in p = 0.001* p = 0.033* p = 0.320* GeneTranscription (normalized to GAPDH) Baseline compared to End of StudyTNF Receptor Superfamily 1A Relative 0.61 ± 0.31 (14) 0.83 ± 0.54 (11)1.00 ± 0.61 (13) 0.145* 0.501* 0.722* 0.124* Quantity 0.57 (0.17 − 1.15)0.74 (0.18 − 1.91) 0.86 (0.35 − 2.67) change in p = 0.001* p = 0.097* p= 0.385* Gene Transcription (normalized to GAPDH) Baseline compared toEnd of Study TNF Receptor Superfamily 1B Relative 0.67 ± 0.38 (14) 0.75± 0.58 (11) 1.04 ± 0.70 (13) 0.217* 0.237* 0.936* 0.353* Quantity 0.58(0.18 − 1.44) 0.58 (0.09 − 1.92) 0.93 (0.21 − 3.1) change in p = 0.004*p = 0.056* p = 0.432* Gene Transcription (normalized to GAPDH) Baselinecompared to End of Study N, number; SD, standard deviation; Min minimum;Max, maximum ^(Δ) Between-group comparisons were made using ANCOVA.^(δ)Pairwise between-group comparisons were made using the Tukeyprocedure. Within-group comparisons were made using the paired Studentt-test. Within-group comparisons were made using the paired Studentt-test. Probability values P ≤ 0.05 are statistically significant.

The muscle biopsies taken on Day 0 and Day 57±3 from participants (n=33)were used to evaluate the effect of Sample 1 and 2 or placebo on proteinexpression/activity of the mTOR/S6K/4EBP1 protein synthesis and NFκβ(p65/p50) pro-inflammatory signaling pathways. Protein was isolated frommuscle tissue collected at baseline and end-of-study, when pre- andpost-exercise biopsies were collected. Post-exercise muscle biopsieswere obtained approximately 1h following the pre-exercise biopsies.Western blotting was done with equal concentrations (40 μg) of proteinseparated on 8-12% SDS-PAGE gels, which were then transferred ontonitrocellulose membranes, and immune-labeled with antibodies specific toeach protein of interest. Detection of these antibodies was done usingenhanced chemiluminescence (ECL).

As shown in Table 9, there were notable changes in the muscle proteinlevels of participants taking Sample 1 and Sample 2 from week 0(baseline) to week 8 (end of study) pre-exercise. In Table 9, in orderto minimize variability between runs, the areas under of the curve werestandardized relative to the micrograms of protein.

FIGS. 6-9 illustrate changes in mTOR expression from baseline to week 8in each treatment group. As seen in Table 9 and FIG. 9, within group,participants on Sample 2 showed a significant 81% increase in total mTORexpression (p=0.017) at week 8-pre-exercise relative to baseline mTORlevels.

FIGS. 14-17 illustrate changes in Phospho-4EBP1 and Total-4EBP1 frombaseline to week 8 in each treatment group. As shown in Table 9 and FIG.15, Sample 2 had a trend towards a 24% reduction in the phosphorylationof 4EBP-1 protein (p=0.057) at week 8—pre-exercise compared to week 0.

FIGS. 18-21 illustrate changes in Phospho-P65 and Total-P65 frombaseline to week 8 in each treatment group. As seen in Table 9 and FIG.22, the Sample 1 group showed a significant within group decrease of 38%for p65 phosphorylation of the NF-κβ complex (p=0.021) when comparingweek 0 to week 8—pre-exercise. As shown in FIG. 21, there was a trendtowards the Sample 1 group having a significantly reduced amount oftotal p65 expression compared to the placebo group when comparing week 0to week 8—pre-exercise (p=0.087).

As shown in Table 9 and FIG. 6, there were also significant within groupchanges in muscle protein levels of participants taking Sample 1 andSample 2 from week 0 (baseline) to week 8 (end of study) post-exercise.As illustrated in FIGS. 6-9, phosphorylation of mTOR was significantlyincreased by 41% (p=0.015) in the Sample 1 group and reduced by 35%(p=0.051) in the Sample 2 group. However, as FIG. 9 shows, total mTORexpression was increased by 56% (p=0.058) in the Sample 2 group, whichwas borderline significant and may compensate for the decrease in itsphosphorylation for this group at week 8—post-exercise relative tobaseline. As seen in FIG. 7, between groups, there was a significantdifference between the Sample 1 and Sample 2 groups when comparing thechanges from week 0 to week 8—post-exercise of phospho-mTOR (p=0.016).

FIGS. 10-13 illustrate changes in Phospho-S6K and Total-S6K frombaseline to week 8 in each treatment group. As seen in FIG. 11, S6Kphosphorylation was significantly decreased by 36% (p=0.019) in theSample 1 group when comparing week 0 to week 8—post-exercise. On theother hand, as shown in FIG. 13, total S6K protein expression wassignificantly increased by 101% (p=0.038) in the Sample 1 group whencomparing week 0 to week 8—post-exercise, and trended towards asignificant increase of 47% in the Sample 2 group (p=0.098) as well.

As illustrated in FIGS. 14 and 15, the phosphorylation of 4EBP-1 wassignificantly decreased by 56% (p=0.04) and 38% (p=0.019), respectivelyin Sample 1 and Sample 2 groups when comparing within group changes atweek 0 to week 8—post-exercise.

As shown in FIG. 19, phosphorylation of the p65 subunit of NF-κβ wassignificantly reduced by 56% (p=0.017) in the Sample 1 group at week8—post-exercise relative to baseline.

FIGS. 22 and 23 illustrate changes in Total-P50 from baseline to week 8in each treatment group. As seen in FIGS. 25 and 26, there were nosignificant changes in total p50 expression from baseline to week 8 inany of the treatment groups.

In FIG. 24, a representative immuno-blot for each indicated protein isshown for each treatment group (at baseline vs. end of study, 8 weeks).GAPDH was used as an internal loading control. In FIG. 14, T=total andP=phospho antibodies.

The subject number (N=33) for Table 9 is reflected by the processing andisolation of protein from subjects muscle biopsies. Baseline and end ofstudy muscle samples were required for this analysis (to see relativechanges in the requested markers) and some of the subjects did not haveenough skeletal muscle to yield the amount of protein required to runthe molecular analysis.

TABLE 9 Quantity of Proteins at Week 0 and Pre and Post Exercise at Week8 for All Participants in the PP Population (N = 33). CarnipureCarnipure Placebo Product 1 Product 2 Mean ±SD (n) Mean ±SD (n) Mean ±SD(n) P Value^(¤) Median Median Median Carnipure (Min − Max) (Min − Max)(Min − Max) Placebo Placebo Product 1 Within Within Within vs vs VsGroup P Group P Group P Over- Carnipure Carnipure Carnipure Value ValueValue all ^(Δ) Product 1^(δ) Product 2^(δ) Product 2^(δ) AUC ofPhospho-mTOR per Microgram Protein (×10¹) Week 0 8.5 ± 4.4 (10) 6.8 ±4.5 (11) 7.6 ± 3.4 (12) — — — — Baseline 6.8 (2.4 − 15.9) 4.5 (2.3 − 17)6.9 (2.4 − 15) Pre- Exercise Week 8 8.5 ± 3.9 (10) 7.9 ± 5.4 (11) 8.7 ±4.2 (12) — — — — Pre- 8 (2.8 − 13.3) 5.9 (2.6 − 19) 9.5 (2.7 − 14.7)Exercise Week 8 7.0 ± 4.8 (10) 9.6 ± 4.6 (11) 5.1 ± 2.1 (11) — — — —Post- 5.8 (1.5 − 15.7) 9.7 (2.4 − 16) 4.4 (2 − 8.8) Exercise Change 0.0± 5.5 (10) 1.1 ± 5.7 (11) 1.1 ± 4.7 (12) 0.978* 1.000* 0.985* 0.980*from 0.7 (−10.5 − 7) 1.5 (−13 − 9.6) 0.4 (−8.3 − 8.8) Week 0 to p =0.938* p = 0.442* p = 0.556* Week 8 Pre- Exercise Change −1.5 ± 3.4 (10)2.8 ± 3.3 (11) −2.7 ± 3.5 (11) 0.016* 0.078* 0.800* 0.016* from −0.9(−9.5 − 3.1) 2.3 (−2.5 − 7.2) −2 (−8.5 − 2.1) Week 0 to p = 0.255* p =0.015* p = 0.051* Week 8 Post- Exercise Change −1.5 ± 5.6 (10) 1.7 ± 5.5(11) −3.4 ± 5.3 (11) 0.067* 0.246* 0.776* 0.061* from −2.8 (−9.8 − 9.8)2.8 (−11.1 − 10.5) 1.2 (−11.8 − 3.9) Pre- p = 0.245* p = 0.237* p =0.105* Exercise to Post- Exercise Week 8 AUC of Total mTOR per MicrogramProtein (×10¹) Week 0 5.6 ± 4.7 (10) 6.0 ± 5.3 (11) 4.3 ± 4.7 (12) — — —— Baseline 5.1 (0.2 − 13.5) 5.9 (0.5 − 17.9) 2.8 (0.7 − 17.7) Pre-Exercise Week 8 5.0 ± 5.2 (10) 7.5 ± 7.0 (11) 7.8 ± 7.2 (12) — — — —Pre- 3.3 (0.1 − 15.9) 6 (0.3 − 18.5) 4.7 (1.4 − 26.1) Exercise Week 86.7 ± 4.4 (10) 7.4 ± 6.4 (11) 7.0 ± 5.5 (11) — — — — Post- 5.7 (1.4 −15.7) 5.6 (1 − 18.7) 4.2 (1.1 − 18.3) Exercise Change −0.7 ± 4.7 (10)1.5 ± 5.5 (11) 3.5 ± 4.3 (12) 0.122* 0.758* 0.112* 0.364* from −0.3(−10.7 − 6.7) 0.5 (−8.5 − 9.8) 2.7 (−0.8 − 10.9) Week 0 to p = 0.590* p= 0.795* p = 0.017* Week 8 Pre- Exercise Change 1.0 ± 3.6 (10) 1.5 ± 5.8(11) 2.4 ± 5.6 (11) 0.858* 0.864* 0.994* 0.905* from 0.8 (−6.6 − 6.5)0.8 (−6.7 − 13.7) 2.1 (−9 − 10.6) Week 0 to p = 0.121* p = 0.487* p =0.058* Week 8 Post- Exercise Change 1.7 ± 4.2 (10) −0.0 ± 7.0 (11) −1.3± 7.5 (11) 0.591* 0.721* 0.587* 0.967* from 1.3 (−7.5 − 7.1) 1.1 (−16.5− 10) −0.3 (−17.3 − 8) Pre- p = 0.057* p = 0.650* p = 0.637* Exercise toPost- Exercise Week 8 AUC of Phospho-S6K per Microgram Protein (×10¹)Week 0 8.3 ± 4.1 (10) 10.8 ± 3.4 (11) 8.4 ± 4.3 (12) — — — — Baseline 8(1.7 − 14.3) 9.7 (8.1 − 18) 8 (1.6 − 17.4) Pre- Exercise Week 8 8.2 ±4.5 (10) 11.4 ± 3.6 (11) 11.2 ± 8.9 (12) — — — — Pre- 8.2 (2.8 − 15.9)11.7 (4.8 − 15.2) 8.4 (3.9 − 35.2) Exercise Week 8 9.4 ± 9.4 (10) 6.9 ±1.8 (11) 7.3 ± 4.6 (11) — — — — Post- 7.1 (1.1 − 34.3) 7.2 (4.9 − 10.6)6 (2 − 17.8) Exercise Change −0.1 ± 4.0 (10) 0.5 ± 4.5 (11) 2.7 ± 7.5(12) 0.449* 0.546* 0.482* 0.999* from 1.4 (−6.2 − 5.3) 0.2 (−9 − 6.5)1.8 (−5.3 − 24.6) Week 0 to p = 0.959* p = 0.816* p = 0.159* Week 8 Pre-Exercise Change 1.1 ± 11.1 (10) −3.9 ± 4.0 (11) −0.8 ± 3.4 (11) 0.864*0.852* 0.972* 0.943* from −0.3 (−8 − 30.8) −2.4 (−13 − −0.9) 0.4 (−9.4 −4) Week 0 to p = 0.774* p = 0.005* p = 0.384* Week 8 Post- ExerciseChange 1.1 ± 10.5 (10) −4.4 ± 4.8 (11) −4.0 ± 7.8 (11) 0.870* 0.946*0.858* 0.977* from −2.8 (−7.8 − 28.9) −3.8 (−9.9 − 5.8) −3.1 (−25.6 −3.4) Pre- p = 0.812* p = 0.019* p = 0.046* Exercise to Post- ExerciseWeek 8 AUC of Total S6K per Microgram Protein (×10¹) Week 0 18.6 ± 14.0(10) 17.7 ± 12.5 (11) 19.2 ± 9.0 (12) — — — — Baseline 16.1 (4.7 − 44)17.3 (0.4 − 41.9) 18.9 (7.5 − 31.7) Pre- Exercise Week 8 26.6 ± 14.7(10) 20.3 ± 12.2 (11) 19.5 ± 7.3 (12) — — — — Pre- 24.1 (1.3 − 57.6) 23(2 − 43) 20.4 (8.1 − 29.8) Exercise Week 8 28.3 ± 28.0 (10) 35.6 ± 37.1(11) 28.3 ± 17.5 (11) — — — — Post- 20.3 (6.4 − 102.4) 18.2 (6.1 −122.5) 28.2 (6.8 − 57) Exercise Change 8.0 ± 13.8 (10) 2.6 ± 13.9 (11)0.2 ± 13.0 (12) 0.825* 0.841* 0.861* 0.998* from 10.8 (−17.8 − 26.9) 1.6(−25.4 − 20.5) −0.1 (−23.5 − 20.1) Week 0 to p = 0.373* p = 0.310* p =0.833* Week 8 Pre- Exercise Change 9.8 ± 29.5 (10) 17.9 ± 34.5 (11) 9.1± 13.3 (11) 0.665* 0.692* 0.996* 0.739* from 1.3 (−24.1 − 83.4) 7 (−11.3− 104.8) 4.3 (−11.7 − 31.5) Week 0 to p = 0.180* p = 0.038* p = 0.098*Week 8 Post- Exercise Change 2 ± 37 (10) 15 ± 37 (11) 10 ± 16 (11)0.919* 0.920* 0.946* 0.997* from −11 (−27 − 101) 4 (−30 − 99) 1 (−12 −35) Pre- p = 0.965* p = 0.162* p = 0.218* Exercise to Post- ExerciseWeek 8 AUC of Phospho-4EBP1 per Microgram Protein (×10¹) Week 0 23.1 ±14.6 (10) 27.5 ± 20.6 (11) 22.9 ± 12.6 (12) — — — — Baseline 21.8 (2 −41.2) 28.4 (4.3 − 64) 23.3 (6.8 − 52.4) Pre- Exercise Week 8 14.6 ± 11.1(10) 21.0 ± 21.8 (11) 17.5 ± 14.6 (12) — — — — Pre- 12.5 (3.9 − 34) 12.8(0.4 − 62) 13.2 (2 − 46) Exercise Week 8 16.2 ± 10.5 (10) 12.2 ± 6.5(11) 13.9 ± 11.9 (11) — — — — Post- 17.3 (3.4 − 31.1) 9.7 (4.8 − 22.9)10.9 (1.8 − 33.9) Exercise Change −8.5 ± 15.4 (10) −6.5 ± 12.7 (11) −5.5± 16.6 (12) 0.924* 0.918* 0.985* 0.968* from −1.3 (−34.8 − 6) −2 (−29.8− 14.3) −4.8 (−39 − 21.5) Week 0 to p = 0.276* p = 0.153* p = 0.057*Week 8 Pre- Exercise Change −6.9 ± 14.2 (10) −15.3 ± 19.8 (11) −8.8 ±11.1 (11) 0.583* 0.802* 0.556* 0.908* from −8 (−35.2 − 11.7) −5.5 (−56.1− 4.3) −4.9 (−25 − 4.2) Week 0 to p = 0.316* p = 0.040* p = 0.019* Week8 Post- Exercise Change 1.6 ± 15.0 (10) −8.8 ± 23.6 (11) −0.9 ± 15.9(11) 0.765* 0.915* 0.745* 0.938* from 2.5 (−28 − 24.7) −0.7 (−54.1 −17.9) −0.1 (−26.2 − 25.2) Pre- p = 0.816* p = 0.898* p = 0.865* Exerciseto Post- Exercise Week 8 AUC of Total 4EBP1 per Microgram Protein (×10¹)Week 0 10.1 ± 5.2 (10) 20.0 ± 10.6 (11) 20.8 ± 11.4 (12) — — — —Baseline 8.3 (4.8 − 19.9) 18.2 (3.5 − 38.8) 18.7 (3.9 − 47.1) Pre-Exercise Week 8 21.1 ± 32.3 (10) 16.4 ± 13.5 (11) 13.8 ± 7.0 (12) — — —— Pre- 10.3 (1.8 − 110.6) 12.4 (5.7 − 42.9) 14.7 (0.8 − 28.2) ExerciseWeek 8 14.6 ± 11.0 (10) 17.8 ± 7.3 (11) 16.1 ± 13.6 (11) — — — — Post-11.7 (1.1 − 34.3) 17.9 (3.8 − 27.9) 12.8 (3.3 − 51.4) Exercise Change11.0 ± 33.1 (10) −3.6 ± 10.9 (11) −7.0 ± 14.3 (12) 0.827* 0.967* 0.821*0.916* from 1.9 (−4.8 − 104.7) −4.3 (−19.7 − 17.6) −2.6 (−38.88 − 10.2)Week 0 to p = 0.619* p = 0.170* p = 0.151* Week 8 Pre- Exercise Change4.5 ± 8.9 (10) −2.2 ± 12.6 (11) −5.0 ± 11.4 (11) 0.713* 0.873* 0.972*0.698* from 2.6 (−7.4 − 21.3) −3 (−23.6 − 17.3) −3.2 (−27.1 − 15.7) Week0 to p = 0.819* p = 0.817* p = 0.114* Week 8 Post- Exercise Change −6.5± 31.4 (10) 1.4 ± 16.5 (11) 3.6 ± 15.5 (11) 0.461* 0.428* 0.773* 0.828*from 3.6 (−92.7 − 19.6) 5 (−30 − 20.5) 0.5 (−10.9 − 43.1) Pre- p =0.725* p = 0.497* p = 0.607* Exercise to Post- Exercise Week 8 AUC ofPhospho-p65 per Microgram Protein (×10¹) Week 0 9.0 ± 10.1 (10) 19.4 ±15.2 (11) 10.9 ± 10.2 (12) — — — — Baseline 5.4 (2.1 − 30.9) 16.1 (4.3 −54.4) 10.3 (2 − 40.3) Pre- Exercise Week 8 10.0 ± 8.9 (10) 12.1 ± 8.2(11) 11.1 ± 10.2 (12) — — — — Pre- 5.3 (1 − 23.8) 9.3 (4.6 − 26.5) 7.9(1.6 − 32.2) Exercise Week 8 7.4 ± 7.0 (10) 8.6 ± 4.3 (11) 7.4 ± 4.1(11) — — — — Post- 8.1 (0.4 − 20.6) 9.5 (1.2 − 18.6) 6.6 (1.2 − 15.4)Exercise Change 1.0 ± 6.8 (10) −7.4 ± 12.0 (11) 0.2 ± 6.6 (11) 0.488*0.508* 0.973* 0.574* from −0.5 (−10.6 − 15) −4 (−37 − 9.8) −0.6 (−8.1 −16.8) Week 0 to p = 0.837* p = 0.021* p = 0.705* Week 8 Pre- ExerciseChange −1.7 ± 10.5 (10) −10.8 ± 15.2 (11) −3.3 ± 10.9 (11) 0.522* 0.737*0.497* 0.946* from −1.6 (−22.8 − 18.5) −6.6 (−48.9 − 5.3) 0.5 (−33.7 −6.2) Week 0 to p = 0.272* p = 0.017* p = 0.444* Week 8 Post- ExerciseChange −2.6 ± 9.5 (10) −3.5 ± 9.0 (11) −3.2 ± 11.3 (11) 0.302* 0.330*0.407* 0.982* from −4.8 (−12.3 − 19.3) 1.5 (−19.8 − 5.4) −0.1 (−25.6 −8.1) Pre- p = 0.303* p = 0.275* p = 0.701* Exercise to Post- ExerciseWeek 8 AUC of Total p65 per Microgram Protein (×10¹) Week 0 13.8 ± 11.8(10) 6.8 ± 5.6 (11) 11.4 ± 8.2 (12) — — — — Baseline 9.2 (1.7 − 31) 3.5(1.4 − 18.2) 10.3 (1 − 26.1) Pre- Exercise Week 8 14.2 ± 9.2 (10) 6.0 ±4.6 (11) 9.0 ± 5.4 (12) — — — — Pre- 13.2 (2.6 − 26.5) 5 (1.5 − 16.7)7.7 (1.5 − 21.1) Exercise Week 8 11.1 ± 8.1 (10) 8.6 ± 7.1 (11) 8.8 ±6.8 (11) — — — — Post- 8.3 (3.1 − 27.5) 6 (1.3 − 21.1) 5.8 (2.9 − 26.6)Exercise Change 0.4 ± 9.1 (10) −0.8 ± 5.9 (11) −2.4 ± 8.4 (12) 0.103*0.087* 0.372* 0.617* from 1.5 (−14.7 − 17.6) 0.1 (−11.4 − 9.1) −0.5(−19.7 − 6) Week 0 to p = 0.245* p = 0.775* p = 0.787* Week 8 Pre-Exercise Change −2.7 ± 13.5 (10) 1.8 ± 7.1 (11) −3.5 ± 9.1 (11) 0.671*0.667* 0.796* 0.969* from −1.9 (−25.2 − 19.3) 2.7 (−6 − 19.7) −2.2(−20.9 − 10.4) Week 0 to p = 0.952* p = 0.565* p = 0.367* Week 8 Post-Exercise Change −3.1 ± 9.4 (10) 2.6 ± 9.4 (11) −0.9 ± 10.5 (11) 0.509*0.478* 0.773* 0.821* from −3.3 (−18.7 − 9.5) 1 (−9.6 − 19.6) −2.6 (−15.3− 23.3) Pre- p = 0.323* p = 0.379* p = 0.787* Exercise to Post- ExerciseWeek 8 AUC of Total-p50 per Microgram Protein (×10¹) Week 0 15.7 ± 6.0(10) 13.9 ± 9.2 (11) 11.6 ± 6.3 (12) — — — — Baseline 16.8 (7.7 − 21.9)9.5 (5.7 − 33.6) 8.5 (3.3 − 21.6) Pre- Exercise Week 8 18.2 ± 6.5 (10)13.8 ± 8.0 (11) 12.8 ± 5.3 (12) — — — — Pre- 19.1 (7.8 − 27.7) 14.3 (6 −34.7) 11.7 (4.4 − 21.6) Exercise Week 8 21.6 ± 14.1 (10) 10.6 ± 6.7 (11)17.4 ± 12.6 (11) — — — — Post- 21.6 (3.2 − 48.4) 9.1 (3.2 − 21.6) 15.5(4.5 − 41.1) Exercise Change 2.5 ± 6.0 (10) −0.0 ± 4.4 (11) 1.2 ± 5.1(12) 0.380* 0.397* 0.487* 0.986* from 1.8 (−9 − 12) 0.5 (−7.1 − 9.6) 0.4(−7.6 − 9.6) Week 0 to p = 0.237* p = 0.599* p = 0.352* Week 8 Pre-Exercise Change 5.8 ± 15.2 (10) −3.2 ± 10.3 (11) 5.4 ± 11.7 (11) 0.209*0.243* 0.967* 0.331* from 1.7 (−12.9 − 37.7) −0.3 (−27.7 − 6.7) 1.2 (−10− 33) Week 0 to p = 0.683* p = 0.294* p = 0.248* Week 8 Post- ExerciseChange 3.3 ± 15.6 (10) −3.2 ± 11.2 (11) 4.2 ± 13.4 (11) 0.174* 0.165*0.778* 0.419* from Pre- 2.1 (−14.5 − 37) 0.9 (−28.8 − 7.1) 4.6 (−11.5 −33.3) Exercise p = 0.912* p = 0.240* p = 0.727* to Post- Exercise Week 8N, number; SD, standard deviation; Min, minimum; Max, maximum; AUC, areaunder the curve ^(Δ) Between-group comparisons were made using ANCOVA.^(δ)Pairwise between-group comparisons were made using the Tukeyprocedure. Within-group comparisons were made using the paired Studentt-test. *Logarithmic transformation required to achieve normality.Probability values P ≤ 0.05 are statistically significant.

Anthropometrics and vital signs were taken for each participant prior toDay 0 and on Day 57±3. As seen in Table 10, there were no significantbetween-group differences for both systolic and diastolic blood pressureas well as heart rate, weight, and BMI for all post randomization timepoints. Within groups, there was a significant change in diastolic bloodpressure for the Sample 1 group, as well as mean heart rate in theSample 2 group relative to baseline. All blood pressure and heart ratevalues were within acceptable clinical range.

TABLE 10 Vital Signs for All Participants Enrolled in the Study atBaseline and Week 8 (N = 42). Placebo Sample 1 Sample 2 Mean ± SD (n)Mean ± SD (n) Mean ± SD (n) Median (Min − Max) Median (Min − Max) Median(Min − Max) Within Group P Within Group P Within Group P P Value ValueValue Value ^(Δ) Mean Systolic Blood Pressure (mmHg) Baseline 112.2 ±9.5 (14)  119.1 ± 10.3 (14) 115.7 ± 12.9 (14) — (Week 0) 112.7(97.3-131.3)    117.5 (104.7-141.7) 115.7 (96-138.3) Visit 3  112.0 ±11.1 (13)  121.2 ± 11.1 (14) 118.3 ± 12.9 (14) — (Week 8) 112.7(94-134.3)   121.3 (101.3-140) 118.7 (94-142)   Change from  0.4 ± 11.0(13)  2.1 ± 8.5 (14)  2.6 ± 12.3 (14) 0.447 Baseline to 0.3 (−18.7-20) 2.7 (−14-21)   1.7 (−18-20) Week 8 p = 0.902 p = 0.373 p = 0.435 MeanDiastolic Blood Pressure (mmHg) Baseline 69.2 ± 6.9 (14)  70.0 ± 7.0(14) 71.9 ± 9.1 (14) — (Week 0) 70.2 (54.7-78.7)   70.8 (60.7-84.7)  73.3 (52.3-85.7) Visit 3 69.3 ± 6.1 (13)  74.1 ± 6.3 (14)  74.9 ± 11.9(14) — (Week 8) 69.3 (60-80)   73.5 (62.7-88)    75.5 (55.3-96.7) Changefrom 0.1 ± 7.4 (13)  4.1 ± 6.0 (14)  3.0 ± 7.6 (14) 0.223 Baseline to−0.3 (−10.7-19.3)   5.8 (−11.3-11.3)   3.5 (−9.7-16.7) Week 8 p = 0.952p = 0.024 p = 0.162 Mean Heart Rate (BPM) Baseline 73.6 ± 12.6 (14) 71.6± 7.3 (14) 71.7 ± 6.9 (14) — (Week 0) 69.8 (52-103.3)    72 (56.3-83.3)  72.5 (54.7-81.3) Visit 3 68.1 ± 7.2 (13)  69.9 ± 7.1 (14) 66.6 ± 5.2(14) — (Week 8) 69.3 (53.3-81.3)  69.2 (56-85)     66.7 (58.7-74.7)Change from −3.2 ± 10.6 (13) −1.7 ± 8.4 (14) −5.1 ± 7.6 (14) 0.391Baseline to −3.3 (−20.7-13.3) −1.3 (−16-13.3) −4.7 (−22.7-8) Week 8 p =0.297 p = 0.464 p = 0.024 Weight (kg) Baseline 72.1 ± 8.3 (14)   74.8 ±13.4 (14)  76.5 ± 14.2 (14) — (Week 0) 71.2 (60.5-86.2)  75.3(55.8-98.2) 72.8 (56-102)  Visit 3 73.1 ± 8.0 (13)   75.3 ± 13.4 (14) 77.0 ± 14.1 (14) — (Week 8) 73 (61.5-88)  75 (57-97.9)  74.5(56.5-102.3) Change from 0.23 ± 0.82 (13)  0.46 ± 1.89 (14)  0.48 ± 1.23(14)  0.678^(¤) Baseline to 0.2 (−1.5-1.8)  0.65 (−3.2-4.6)  0.5(−2.2-3) Week 8  p = 0.294^(‡)  p = 0.330^(‡)  p = 0.116^(‡) BMI (kg/m²)Baseline 26.00 ± 1.83 (14)  26.54 ± 3.03 (14) 27.60 ± 3.05 (14) — (Week0) 25.16 (23.49-30.18) 26.78 (21.8-31.56)   27.31 (22.58-32.48) Visit 326.22 ± 2.00 (13)  26.70 ± 2.96 (14) 27.77 ± 2.89 (14) — (Week 8)  25.6(23.67-30.81)  26.89 (22.27-31.46)   27.69 (22.66-32.58) Change from0.10 ± 0.28 (13)  0.16 ± 0.66 (14)  0.17 ± 0.47 (14) 0.820 Baseline to0.07 (−0.45-0.63)  0.24 (−1.17-1.51)   0.18 (−0.88-1.13) Week 8 p =0.239 p = 0.372 p = 0.195 N, number; SD, standard deviation; Min,minimum; Max, maximum; mmHg, millimeters of mercury; BPM, beats perminute. ^(Δ) Between-group comparisons were made using ANCOVA.^(¤)Between group were made using the Kruskal Wallis test. Within-groupcomparisons were made using the paired Student Hest. ^(‡)Within groupwere made using the Sign Rank test. Probability values P ≥ 0.05 arestatistically significant.

Blood samples were taken for each participant taken prior to Day 0 andon Day 57±3 to measure hematology and clinical chemistry parameters.Table 11 demonstrates that there were no significant between groupchanges in hematology and clinical chemistry parameters forparticipants, with the exception of creatinine (P=0.027), estimatedglobular filtration rate (P=0.029), and aspartate transaminase (P=0.034)concentrations. However, these markers remained within their establishedclinical reference ranges.

There were five safety parameters that showed significant within groupchange from screening to the end of study (week 8). White blood cellcount (P=0.021), lymphocyte count (P=0.022), and aspartate transaminaselevels (P=0.033) increased for subjects on Sample 1, while chloridelevels decreased significantly (P=0.013) for this group. The creatinineconcentration (P=0.05) was significantly increased for the Sample 2group. Estimated globular filtration rate (P=0.069) was notsignificantly changed for subjects within the Sample 2 group. Overall,all hematological and clinical parameters were within acceptableclinical range for this population.

TABLE 11 Haematology and Clinical Chemistry Parameters for All EnrolledParticipants at Screening and Week 8 (N = 42). Placebo Sample 1 Sample 2Mean ± SD (n) Mean ± SD (n) Mean ± SD (n) Median (Min-Max) Median(Min-Max) Median (Min-Max) P Within Group P Value Within Group P ValueWithin Group P Value Value ^(Δ) Hemoglobin Concentration (g/L) Screening138.4 ± 7.9 (14) 144.0 ± 10.0 (14) 140.3 ± 12.6 (14) — 137.5 (128-156)143.5 (131-159) 136 (124-167) Visit 3 138.6 ± 6.5 (14) 141.6 ± 9.3 (13)137.9 ± 12.2 (14) — (Week 8)   138 (128-151)   140 (124-154) 138(116-159) Change  0.3 ± 7.6 (14)  −1.2 ± 6.5 (13)  −2.4 ± 7.1 (14)0.559^(¤) from  1.5 (−15-11)    1 (−14-12)  0 (−18-7) Screening p =0.834^(‡) p = 0.505^(‡) p = 0.307^(‡) to Week 8 Hematocrit (L/L)Screening 0.4021 ± 0.0236 (14)   0.4229 ± 0.0246 (14)   0.4093 ± 0.0300(14) —  0.4 (0.37-0.45)   0.43 (0.39-0.46)  0.4 (0.37-0.47) Visit 30.4064 ± 0.0178 (14)   0.4162 ± 0.0260 (13)   0.4036 ± 0.0310 (14) —(Week 8)  0.41 (0.38-0.44)   0.41 (0.37-0.46) 0.405 (0.34-0.45) Change0.0043 ± 0.0217 (14) −0.0046 ± 0.0176 (13) −0.0057 ± 0.0187 (14)0.374^(¤) from 0.005 (−0.04-0.04) −0.01 (−0.04-0.03)    0 (−0.05-0.02)Screening p = 0.420^(‡) p = 0.337^(‡) p = 0.262^(‡) to Week 8 WhiteBlood Cell Count (×E9/L) Screening   5.91 ± 1.16 (14) 5.88 ± 1.88 (14)  6.25 ± 1.86 (14) — 5.65 (4.6-7.9) 5.3 (3.8-9.3)   6 (3.8-10.8) Visit 3  5.64 ± 1.32 (14) 6.59 ± 1.75 (13)   6.17 ± 1.91 (14) — (Week 8)  5.6(3.7-8.7) 6.4 (4.3-10.5) 6.15 (3.9-11) Change −0.28 ± 1.10 (14) 0.68 ±0.83 (13) −0.08 ± 0.87 (14) 0.054^(¤) from −0.3 (−2.2-1.3) 0.5(−0.4-2.3)   0 (−1.4-1.2) Screening p = 0.441^(‡) p = 0.021^(‡) p =0.753‡ to Week 8 Red Blood Cell Count (×E12/L) Screening 4.48 ± 0.26(14)   4.72 ± 0.40 (14)   4.55 ± 0.35 (14) — 4.47 (3.94-5.04)    4.7(4.01-5.34)   4.49 (4.08-5.12) Visit 3 4.58 ± 0.22 (14)   4.66 ± 0.44(13)   4.44 ± 0.34 (14) — (Week 8) 4.56 (4.23-4.95)   4.61 (3.96-5.4)  4.42 (3.87-5.01) Change 0.09 ± 0.35 (14) −0.07 ± 0.21 (13) −0.11 ±0.30 (14) 0.222* from 0.03 (−0.41-0.93) −0.05 (−0.48-0.31) −0.07(−0.97-0.21) Screening p = 0.329* p = 0.244* p = 0.212* to Week 8 MeanCorpuscular Volume (fL) Screening   89.9 ± 3.0 (14) 89.7 ± 4.6 (14) 89.9± 3.5 (14) —     89 (85.1-96.3) 88.2 (83.3-100.2) 90.8 (81.2-93.6) Visit3   88.6 ± 3.7 (14) 89.2 ± 3.0 (13) 90.8 ± 3.0 (14) — (Week 8)     89(80.5-96.3)   89 (84.6-93.8) 91.2 (84.4-95.3) Change −1.28 ± 3.55 (14)0.40 ± 1.16 (13) 0.86 ± 2.21 (14) 0.091^(¤) from −0.15 (−12.6-1.4)  0.8(−3-1.5)  0.4 (−1.9-7.2) Screening p = 0.221^(‡) p = 0.077^(‡) p =0.116^(‡) to Week 8 Mean Corpuscular Hemoglobin (pg) Screening 30.89 ±1.08 (14) 30.61 ± 1.75 (14) 30.86 ± 1.40 (14) —  30.9 (28.9-33) 30.15(28.3-35.2) 31.15 (27.7-33.3) Visit 3 30.32 ± 1.40 (14) 30.45 ± 1.10(13) 31.04 ± 1.25 (14) — (Week 8) 30.35 (27.3-33.2)  30.4 (28.5-32.1)31.35 (28.7-33.6) Change −0.56 ± 1.42 (14)  0.19 ± 0.36 (13)  0.19 ±0.74 (14) 0.093* from −0.15 (−5.2-0.4)  0.2 (−0.6-0.8)  0.1 (−0.5-2.3)Screening p = 0.162* p = 0.070* p = 0.371* to Week 8 Mean CorpuscularHemoglobin Concentration (g/L) Screening 343.6 ± 5.6 (14) 341.1 ± 5.6(14) 342.9 ± 6.7 (14) — 342.5 (336-356) 340 (331-351) 342.5 (333-356)Visit 3 342.2 ± 5.5 (14) 341.2 ± 5.4 (13) 342.1 ± 6.0 (14) — (Week 8)  341 (333-353) 341 (331-349)   340 (335-353) Change  −1.4 ± 6.2 (14) 0.9 ± 3.9 (13)  −0.9 ± 5.1 (14) 0.449^(¤) from   −3 (−11-11)  1 (−7-9) −1.5 (−9-6) Screening p = 0.362^(‡) p = 0.331^(‡) p = 0.599^(‡) to Week8 Red Cell Distribution Width (%) Screening 13.61 ± 0.40 (14) 13.91 ±0.47 (14) 13.58 ± 0.51 (14) — 13.45 (13.1-14.4) 13.9 (12.8-14.5) 13.55(12.8-14.4) Visit 3 13.56 ± 0.34 (14) 13.83 ± 0.48 (13) 13.61 ± 0.56(14) — (Week 8)  13.6 (13-14.3) 13.7 (13.1-14.7) 13.75 (12.3-14.4)Change −0.06 ± 0.30 (14) −0.10 ± 0.43 (13)  0.03 ± 0.41 (14) 0.838 from−0.05 (−0.9-0.4)   0 (−1.2-0.6)  0.15 (−0.9-0.6) Screening p = 0.778* p= 0.123* p = 0.685* to Week 8 Platelet Count (×E9/L) Screening  239 ± 34(14) 265 ± 63 (14)   271 ± 65 (14) — 246 (186-305) 254 (190-424) 268(145-371) Visit 3  235 ± 29 (14) 275 ± 68 (13)   258 ± 63 (14) — (Week8) 238 (190-273) 266 (171-430) 262 (140-362) Change −4.6 ± 20.9 (14) 5.4 ± 15.4 (13) −13.0 ± 28.3 (14) 0.118^(¤) from  3 (−36-42)  5(−19-33)  −5 (−67-41) Screening p = 0.615^(‡) p = 0.289^(‡) p =0.090^(‡) to Week 8 Neutrophil Count (×E9/L) Screening   3.41 ± 0.64(14) 3.17 ± 1.36 (14)   3.59 ± 1.21 (14) —    3.3 (2.4-4.5) 2.95(1.6-6.4) 3.25 (2-6.4) Visit 3   3.15 ± 0.77 (14) 3.63 ± 1.32 (13)  3.34 ± 1.35 (14) — (Week 8)   3.15 (1.6-4.8)  3.7 (1.5-6) 3.15(1.6-6.8) Change −0.26 ± 0.72 (14) 0.44 ± 0.89 (13) −0.25 ± 0.89 (14)0.104^(¤) from −0.55 (−1.1-1.1)  0.2 (−0.8-1.9) −0.2 (−2.4-1) Screeningp = 0.184^(‡) p = 0.146^(‡) p = 0.421^(‡) to Week 8 Lymphocyte Count(×E9/L) Screening   1.84 ± 0.58 (14) 1.91 ± 0.59 (14) 1.96 ± 0.76 (14) —1.95 (0.8-2.9) 1.7 (1.3-3)     2 (0.6-3.5) Visit 3   1.84 ± 0.56 (14)2.13 ± 0.65 (13) 2.03 ± 0.97 (14) — (Week 8) 1.75 (0.9-3)   2 (1.2-3.3)   1.8 (0.6-4.1) Change −0.00 ± 0.53 (14) 0.18 ± 0.23 (13) 0.07 ± 0.95(14) 0.208^(¤) from 0.05 (−1.1-0.9) 0.2 (−0.3-0.5) −0.05 (−0.8-3.1)Screening p = 0.944^(‡) p = 0.022^(‡) p = 0.555^(‡) to Week 8 MonocyteCount (×E9/L) Screening   0.457 ± 0.165 (14) 0.564 ± 0.165 (14) 0.514 ±0.141 (14) — 0.4 (0.3-0.9) 0.55 (0.3-0.9)  0.5 (0.3-0.7) Visit 3   0.450± 0.170 (14) 0.600 ± 0.153 (13) 0.564 ± 0.128 (14) — (Week 8) 0.4(0.3-0.8)  0.6 (0.4-1) 0.55 (0.3-0.7) Change −0.007 ± 0.138 (14) 0.031 ±0.111 (13) 0.050 ± 0.151 (14) 0.732^(¤) from   0 (−0.3-0.2)   0(−0.1-0.3)   0 (−0.2-0.4) Screening p = 0.928^(‡) p = 0.518^(‡) p =0.276^(‡) to Week 8 Eosinophil Count (×E9/L) Screening   0.164 ± 0.101(14)   0.186 ± 0.196 (14) 0.171 ± 0.114 (14) — 0.15 (0-0.4) 0.1(0.1-0.8)  0.1 (0.1-0.5) Visit 3   0.164 ± 0.108 (14)   0.169 ± 0.063(13) 0.186 ± 0.146 (14) — (Week 8)  0.1 (0-0.4) 0.2 (0.1-0.3) 0.15(0-0.6) Change −0.000 ± 0.068 (14) −0.000 ± 0.191 (13) 0.014 ± 0.077(14) 0.405^(¤) from   0 (−0.1-0.1)   0 (−0.6-0.2)   0 (−0.1-0.1)Screening p = 0.824^(‡) p = 0.388^(‡) p = 0.774^(‡) to Week 8 BasophilCount (×E9/L) Screening 0.007 ± 0.027 (14) 0.036 ± 0.050 (14) 0.036 ±0.050 (14) — 0 (0-0.1) 0 (0-0.1) 0 (0-0.1) Visit 3 0.007 ± 0.027 (14)0.038 ± 0.051 (13) 0.036 ± 0.063 (14) — (Week 8) 0 (0-0.1) 0 (0-0.1) 0(0-0.2) Change 0.000 ± 0.000 (14) 0.008 ± 0.028 (13) 0.000 ± 0.078 (14)0.626^(¤) from 0 (0-0) 0 (0-0.1) 0 (−0.1-0.2) Screening p = 1.000^(‡) p= 1.000^(‡) p = 1.000^(‡) to Week 8 Fasting Glucose Concentration(mmol/L) Screening   5.50 ± 0.33 (14) 5.20 ± 0.44 (14)   5.33 ± 0.53(14) — 5.45 (5-6.1) 5.15 (4.6-6)    5.2 (4.6-6.2) Visit 3   5.37 ± 0.56(14) 5.28 ± 0.70 (14)   5.06 ± 0.76 (14) — (Week 8)  5.3 (4.6-6.9)  5.1(4.3-7.1)   5.05 (2.7-5.8) Change −0.13 ± 0.47 (14) 0.08 ± 0.68 (14)−0.27 ± 0.79 (14) 0.762^(¤) from −0.1 (−1.2-0.8)   0 (−0.8-1.9) −0.15(−2.5-0.6) Screening p = 0.344^(‡) p = 0.969^(‡) p = 0.209^(‡) to Week 8Creatinine Concentration (μmol/L) Screening 71.7 ± 13.8 (14) 72.3 ± 11.1(14) 63.8 ± 7.8 (14) —   74 (49-90)    73 (53-88) 64.5 (49-77) Visit 373.1 ± 13.1 (14) 71.6 ± 11.4 (14) 70.7 ± 17.8 (14) — (Week 8) 72.5(53-92)    69 (53-95) 70.5 (33-105) Change  1.4 ± 7.7 (14)^(a, b) −0.7 ±7.4 (14)^(a)  6.9 ± 15.5 (14)^(b) 0.027^(¤) from  1.5 (−21-12) −0.5(−17-15)   5 (−34-34) Screening p = 0.161^(‡) p = 0.623^(‡) p =0.050^(‡) to Week 8 Estimated Globular Filtration Rate (mL/min/1.73 m²)Screening 84.1 ± 17.9 (14) 81.0 ± 10.4 (14) 91.5 ± 15.8 (14) —   81(59-113) 80.5 (64-103) 87.5 (69-119) Visit 3 81.4 ± 15.4 (14) 83.6 ±11.1 (14) 83.8 ± 15.7 (14) — (Week 8) 84.5 (54-102) 78.5 (67-105)   81(62-121) Change −2.8 ± 15.0 (14)^(a, b)  2.6 ± 11.0 (14)^(b) −7.7 ± 18.9(14)^(a) 0.029^(¤) from   −3 (−30-34)   4 (−26-24) −9.5 (−38-43)Screening p = 0.345^(‡) p = 0.147^(‡) p = 0.069^(‡) to Week 8 SodiumConcentration (mmol/L) Screening 144.36 ± 2.37 (14) 144.57 ± 2.50 (14)142.79 ± 2.49 (14) — 144 (141-148) 145 (141-150) 143.5 (138-146) Visit 3145.36 ± 2.17 (14) 144.64 ± 3.18 (14) 142.86 ± 2.28 (14) — (Week 8) 145(142-150) 145 (140-152)   143 (140-148) Change  1.00 ± 2.60 (14)  0.07 ±3.00 (14)  0.07 ± 3.38 (14) 0.549^(¤) from  1.5 (−2-5)  −1 (−3-7)    0(−5-9) Screening p = 0.163^(‡) p = 0.944^(‡) p = 0.918^(‡) to Week 8Potassium Concentration (mmol/L) Screening   5.10 ± 0.47 (14)   5.11 ±0.29 (14)   4.85 ± 0.46 (14) —   5.15 (4-5.7)   5.1 (4.8-5.6)   4.9(3.8-5.7) Visit 3   5.04 ± 0.59 (14)   5.04 ± 0.38 (14)   4.75 ± 0.57(13) — (Week 8)     5 (4-6.4)     5 (4.5-5.6)   4.8 (3.6-5.9) Change−0.06 ± 0.67 (14) −0.08 ± 0.46 (14) −0.08 ± 0.49 (13) 0.599 from −0.05(−1.3-1) −0.1 (−1-0.6) −0.1 (−1.1-0.6) Screening p = 0.861^(‡) p =0.637^(‡) p = 0.806^(‡) to Week 8 Chloride Concentration (mmol/L)Screening 107.64 ± 1.86 (14) 108.29 ± 1.77 (14) 105.50 ± 1.91 (14) — 107(105-112) 109 (105-111) 105.5 (103-108) Visit 3 106.86 ± 2.35 (14)106.50 ± 2.59 (14) 105.00 ± 2.04 (14) — (Week 8) 106 (103-110) 106(101-110)   106 (102-108) Change  −0.79 ± 1.85 (14)  −1.79 ± 2.04 (14) −0.50 ± 2.24 (14) 0.241^(¤) from  −1 (−3-3)  −2 (−5-2)   −1 (−5-4)Screening p = 0.152^(‡) p = 0.013^(‡) p = 0.393^(‡) to Week 8 TotalBilirubin (μmol/L) Screening  9.4 ± 2.7 (14) 10.2 ± 3.0 (14) 10.9 ± 3.5(14) —   9 (6-15)   9.5 (7-19)   11 (6-18) Visit 3 10.4 ± 4.3 (14)  8.8± 2.4 (14) 11.4 ± 3.3 (14) — (Week 8)   9 (5-23)     8 (7-15) 11.5(7-18) Change  1.0 ± 4.1 (14) −1.4 ± 3.8 (14)  0.5 ± 3.2 (14) 0.315^(¤)from 0.5 (−3-14) −0.5 (−12-4)   1 (−3-7) Screening p = 0.671^(‡) p =0.219^(‡) p = 0.776^(‡) to Week 8 Aspartate Transaminase (U/L) Screening24.3 ± 4.5 (14) 23.0 ± 3.6 (14) 26.1 ± 5.5 (14) —  24 (16-34)  22(18-29) 27 (17-33) Visit 3 24.3 ± 3.8 (14) 27.1 ± 7.4 (14) 26.5 ± 10.1(13) — (Week 8)  24 (18-31)  25 (20-48) 24 (17-57) Change  0.0 ± 4.1(14)^(a)  4.1 ± 6.7 (14)^(b)  0.9 ± 11.2 (13)^(a) 0.034^(¤) from 0.5(−11-6) 2.5 (−4-23) −1 (−9-36) Screening p = 0.506^(‡) p = 0.033^(‡) p =0.503^(‡) to Week 8 Alanine Transaminase (U/L) Screening 24.9 ± 9.8 (14)23.1 ± 5.9 (14) 25.5 ± 7.7 (14) — 22.5 (12-47)   22 (15-35)    26(13-38) Visit 3 24.3 ± 5.6 (14) 25.9 ± 7.3 (14) 25.0 ± 12.2 (14) — (Week8)   25 (15-35) 24.5 (14-41)    22 (14-63) Change −0.6 ± 7.0 (14)  2.9 ±6.2 (14) −0.5 ± 14.0 (14) 0.566* from   1 (−17-6)   4 (−7-15) −1.5(−17-43) Screening p = 0.753^(‡) p = 0.150^(‡) p = 0.223^(‡) to Week 8Gamma-Glutamyltransferase (U/L) Screening 24.3 ± 12.9 (14) 19.2 ± 10.2(14) 23.5 ± 14.0 (14) — 19.5 (10-48)  16 (9-50)    19 (9-61) Visit 323.6 ± 10.0 (14) 20.8 ± 15.9 (14) 21.7 ± 14.5 (14) — (Week 8) 20.5(9-44)  14 (10-69)    18 (9-58) Change −0.7 ± 12.9 (14)  1.6 ± 6.4 (14)−1.8 ± 4.6 (14) 0.281^(¤) from   −2 (−38-16) 0.5 (−5-19) −2.5 (−7-11)Screening p = 0.801^(‡) p = 0.779^(‡) p = 0.074^(‡) to Week 8 N, number;SD, standard deviation; Min, minimum; Max, maximum; g, gram; L, liter;fL, femtoliter; pg, picogram; mmol, millimoles; μmol, micromoles; mL,milliliter; min, minutes; m, meters; U, units; μg, microgram; nmol,nanomoles. ^(Δ) Between-group comparisons were made using ANCOVA.Within-group comparisons were made using the paired Student t-test.*Logarithmic transformation was required to achieve normality.^(¤)Between-group comparisons were made using the Kruskal Wallis test.^(‡)Within group were made using the Wilcoxon signed-rank test.Treatment groups with differing letter superscripts are significantlydifferent. Probability values P ≤ 0.05 are statistically significant.

A model was run on creatinine concentration with compliance, treatmentgroup and their interaction as covariates. Table 12 reveals thatcompliance was not related to creatinine concentration.

TABLE 12 Clinical Creatinine Parameters as a Function of Compliance forPP Participants at Screening and Week 8 (N = 39). CreatinineConcentration (μmol/L) Screening 71.7 ± 13.8 (14) 69.5 ± 10.3 (11) 63.8± 7.8 (14) —   74 (49-90) 70 (53-84) 64.5 (49-77) Visit 3 73.1 ± 13.1(14) 69.5 ± 7.6 (11) 70.7 ± 17.8 (14) — (Week 8) 72.5 (53-92) 68 (53-81)70.5 (33-105) Change  1.4 ± 7.7 (14)^(a, b)  0.0 ± 6.1 (11)^(a)  6.9 ±15.5 (14)^(b) 0.030^(¤) from  1.5 (−21-12)  0 (−7-15)   5 (−34-34)Screening p = 0.161^(‡) p = 0.572^(‡) p = 0.050^(‡) to Week 8

The primary endpoint was a composite endpoint of the changes in LeanBody Mass (as assessed by DXA scan) and functional muscle strength (asassessed by 6 Minute Walk Test, Lower Body Dynamometry and Upper BodyDynamometry) of subjects administered Sample 2 compared to thoseadministered placebo from baseline to week 8. The composite endpoint wascalculated by multiplying the results of each component score (kg leanmuscle mass×meters walked×kg resistance lower body×kg resistance upperbody) to derive a composite score (expressed in arbitrary units). Thepercent change in composite score between baseline and week 8 wasderived for comparison between Sample 2 and Placebo.

TABLE 13 Absolute and Percentage Change in the Composite Endpoint atBaseline and at End of the Study for All Participants in the PPPopulation (N = 39). Placebo Sample 1 Sample 2 Mean ± SD (n) Mean ± SD(n) Mean ± SD (n) Between Group P Values Median (Min − Median (Min −Median (Min − Placebo Placebo Sample 1 Max) Max) Max) vs vs vs WithinGroup P Within Group Within Group P Over- Sample Sample Sample Value PValue Value all ^(Δ) 1^(δ) 2^(δ) 2^(δ) Composite Endpoint (×10³)Baseline 11,032 ± 11,501 (14) 10,921 ± 11,142 (11) 6,313 ± 5,485 (14) —— — — (Week 0) 7,870 (559 − 39,977) 5,997 (1,269 − 37,560) 4,466 (677 −19,276) Visit 3 7,611 ± 4,893 (14) 10,620 ± 11,893 (11) 8,367 ± 7,025(14) — — — — (Week 8) 6,334 (360 − 16,803) 6,734 (1,404 − 42,706) 6,475(1,135 − 26,858) Change −3,421 ± 8,073 (14)^(a) −301 ± 3,467 (11)^(a,b)2,054 ± 2,359 (14)^(b) 0.022* 0.826* 0.021* 0.113* from −1,231 (−26,672− 4,814) −374 (−8,681 − 5,146) 858 (−406 − 7,582) Baseline p = 0.232* p= 0.534* p = 0.008* to Week 8 N, number; SD, standard deviation; Min,minimum; Max, maximum; BMI, body mass index; kg, kilogram; m, meter; cm,centimeter ^(Δ) Between-group comparisons were made using ANCOVA.^(δ)Pairwise between-group comparisons were made using the Tukeyprocedure. Within-group comparisons were made using the paired Studentt-test. *Logarihmic transformation was required to achieve normality.^(¤)Between-group comparisons were made using the Kruskal Wallis test.^(‡)Within-group comparisons were made using the signed-rank test.Treatment groups with differing letter superscripts are significantlydifferent. Probability values P ≤0.05 are statistically significant.

As revealed in FIG. 25 and Table 11, there was a significant absolutechange (P=0.008) in the primary composite endpoint [MM (kg)×US (kg)×LS(kg)×6W (m)] for participants taking Sample 2 (n=42, ITT population).This absolute change was significant compared to the placebo group(P=0.016) and approached significance when compared to the Sample 1group (P=0.077). Notably, when expressed as a percentage, the absolutechange in composite endpoints for the Sample 2 group translated into a63.5 percentage point increase over placebo.

Example No. 2

As described above, in one embodiment, the protein building compositionof the present disclosure may be combined with a stabilizer package forimproving one or more properties of the composition. The followingexample provides exemplary formulations, but is not intended to limitthe invention. In fact, the stabilizer package and the process forincorporating the stabilizer package into the composition may be used inany suitable pharmaceutical composition and may not be limited solely tothe protein building composition of the present disclosure.

In one embodiment, the following components may be first mixed together.The components include a protein building composition in accordance withthe present disclosure combined with a polymer binder. In Sample No. 1,the polymer binder is modified starch. In Sample No. 2, the polymerbinder is larch arabinogalactan. The below components can be spray driedinto a granule to form a granular product. For instance, the spraypressure can be about 2 bar while the spray rate can be from about 10 toabout 20 grams per minute.

Sample No. 1 Sample No. 2 (incl. (incl. (dry) moisture) (dry) moisture)Quant. Assay Product Product Quant. Assay Product Product g % g % g % g% Water 1900  0.0% 5  0.50% 1700  0.0% 5  0.50% L-Carnitine 185  99.9%185 18.46%  185  99.9% 185 18.46% Creatine monohydrate 420  87.9% 36936.92%  420  87.9% 369 36.92% L-Leucine 246 100.0% 246 24.62%  246100.0% 246 24.62% vitamin D3 0.10 100.0% 0  0.01%     0.10 100.0% 0 0.01% Sodium stearate/citric acid 10 100.0% 10  1.00%  20 100.0% 20 2.00% esters/rapeseed lecithin Modified Starch 195  95.0% 185 18.50%  0  95.0% 0  0.00% Larch arabinogalactan 0  94.7% 0  0.00%  195  94.7%185 18.50% 0 100.0% 0  0.00%   0 100.0% 0  0.00% Isolated product [g]560.00 1′400.00 800  2′000.00 Properties (granulate, granulate granulategranulate granulate agglom.) Particlesize d10 [μm] 204.59   235.73136.86    235.96 Particlesize d50 [μm] 300.39   331.22 226.10    262.65Particlesize d90 [μm] 450.04   472.33 411.51    363.10 Bulk density[kg/L] 0.56    0.52 0.58     0.68 Moisture [%] (100° C./ 2.30    2.184.28     3.90 25 min) Moisture [%] (KFT) 0.93    0.86 1.88     2.55

After the above granular product is produced, the product can be mixedwith a stabilizer package. The stabilizer package may comprise a dry mixof calcium stearate and silica. The calcium stearate can be added in anamount of about 2.5% by weight, while the silica may be added in anamount of about 0.2% by weight. Because the stabilizer package is a drymix, a pharmaceutical composition is produced that comprises a granularmixture. The stearate and silica serve to stabilize the granularparticles.

To the granular mixture, a fat coating material can be applied. The fatcoating material may comprise a combination of hydrogenated palm oil andpalm stearine. In one embodiment, hydrogenated palm oil is added in anamount of 16% by weight, while palm stearine is added in an amount of 4%by weight of the resulting pharmaceutical composition. In an alternativeembodiment, the hydrogenated palm oil is added in an amount of 24% byweight, while the palm stearine is added in an amount of 6% by weight ofthe pharmaceutical composition.

These and other modifications and variations to the present inventionmay be practiced by those of ordinary skill in the art, withoutdeparting from the spirit and scope of the present invention, which ismore particularly set forth in the appended claims. In addition, itshould be understood that aspects of the various embodiments may beinterchanged both in whole or in part. Furthermore, those of ordinaryskill in the art will appreciate that the foregoing description is byway of example only, and is not intended to limit the invention sofurther described in such appended claims.

1. A method for increasing muscle protein synthesis in healthy mammals,the method comprising administering to the mammal an effective amount ofa protein building composition, said protein building compositioncomprising an amino acid derivative comprising carnitine, combined with,a nitrogenous organic acid or derivative thereof wherein the carnitinecomprises carnitine, derivatives of carnitine, and/or salts thereof,wherein the carnitine is present in the protein building composition ina concentration greater than about 45% by mass; and wherein thenitrogenous organic acid or derivative thereof is present in the proteinbuilding composition at a concentration greater than about 15% by mass.2. (canceled)
 3. (canceled)
 4. A method as defined in claim 1, whereinthe amino acid derivative comprises L-carnitine.
 5. A method as definedin claim 1, wherein the nitrogenous organic acid comprises creatine. 6.A method as defined in claim 5, wherein the nitrogenous organic acidcomprises magnesium chelated creatine.
 7. A method as defined in claim1, wherein the protein building composition further comprises magnesiumor salts thereof.
 8. A method as defined in claim 1, wherein the proteinbuilding composition further comprises vitamin D.
 9. (canceled) 10.(canceled)
 11. A method as defined in claim 1, wherein the proteinbuilding composition is contained in a food product or beverage. 12.(canceled)
 13. A method as defined in claim 1, wherein the proteinbuilding composition is administered at least every one to three days.14. (canceled)
 15. A method as defined in claim 1, wherein the proteinbuilding composition is administered one to four times a day.
 16. Amethod as defined in claim 4, wherein L-carnitine is administered to themammal in an amount from 50 milligrams to 5,000 milligrams per dose. 17.A method as defined in claim 1, further comprising leucine, wherein theleucine is administered to the mammal in an amount from 5 milligrams to5,000 milligrams per dose. 18-19. (canceled)
 20. A method as defined inclaim 1, wherein the mammal is a human. 21.-24. (canceled)
 25. A methodas defined in claim 4, wherein L-carnitine is administered to the mammalin an amount from 50 milligrams to 5,000 milligrams per dose.
 26. Amethod as defined in claim 3, wherein leucine is administered to themammal in an amount from 100 milligrams to 4,000 milligrams per dose.27. A method as defined in claim 5, wherein creatine is administered tothe mammal in an amount from 50 milligrams to 5,000 milligrams per dose.28-29. (canceled)
 30. A method as defined in claim 1, wherein theprotein building composition is administered to the mammal in an amountsufficient to increase lean muscle mass in the mammal.
 31. (canceled)32. A method as defined in claim 1, wherein the protein buildingcomposition is administered to the mammal in an amount sufficient todecrease the amount of TNF-α in the muscles.
 33. A method as defined inclaim 1, wherein the protein building composition is administered to themammal in an amount sufficient to increase mTOR expression in themuscles.
 34. A method as defined in claim 1, wherein mTOR expression isincreased by greater than 40% after activity in comparison to the samemammal that has not received the protein building composition. 35.-80.(canceled)
 81. A method for increasing skeletal muscle protein synthesisin healthy mammals, the method comprising administering to the healthymammal a protein building composition, said protein building compositionconsisting of: an amino acid derivative, wherein the amino acidderivative is L-carnitine, derivatives of L-carnitine, and/or saltsthereof, and a nitrogenous organic acid, wherein the nitrogenous organicacid is creatine, derivatives and/or analogs of creatine, or saltsthereof, and optionally vitamin D and/or at least one excipient, whereinthe carnitine is present in the protein building composition in aconcentration greater than about 45% by mass; and wherein thenitrogenous organic acid or derivative thereof is present in the proteinbuilding composition at a concentration greater than about 15% by mass.82. The method of claim 81, wherein the at least one excipient comprisesan antiadherent, a binder, a synthetic polymer, a coating, a coloringagent, a disintegrant, a filler, a flavoring agent, a glidant, alubricant, a preservative, a sorbent, a sweetener, a vehicle, orcombinations thereof.
 83. The method of claim 81, wherein the amino acidderivative is administered to the healthy mammal in an amount from 50milligrams to 5,000 milligrams per dose, and the nitrogenous organicacid is administered to the healthy mammal in an amount from 50milligrams to 5,000 milligrams per dose.
 84. The method of claim 81,wherein the weight ratio of the carnitine to the creatine is betweenabout 1:1 and about 5:1.
 85. The method of claim 1, wherein the weightratio of the carnitine to the creatine is between about 1:1 and about5:1.
 86. A composition for increasing muscle protein synthesis inhealthy mammals, the composition comprising an amino acid derivativecomprising carnitine, combined with a nitrogenous organic acid orderivative thereof wherein the carnitine comprises carnitine,derivatives of carnitine, and/or salts thereof; wherein the carnitine ispresent in the protein building composition in a concentration greaterthan about 45% by mass; and wherein the nitrogenous organic acid orderivative thereof is present in the protein building composition at aconcentration greater than about 15% by mass.
 87. The composition ofclaim 86, wherein the weight ratio of the carnitine to the creatine isbetween about 1:1 and about 5:1.